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00:01 | Yeah. Okay. Alright folks. . So today, see I sent |
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00:18 | announcements out this morning. Uh Some the casa schedules open now so you |
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00:25 | up for a slot uh see blackboard . So this week uh remember that |
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00:33 | to blackboard quiz types the weekly what's called the weekly quiz. Which |
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00:39 | basically that week's material than the unit quiz precedes the exam. So unique |
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00:46 | gonna be more longer, more more more stuff. Um So checkers 13 |
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00:54 | four will be covered in that not single topic but you know kind of |
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00:58 | decent cross section from those three Um So something on the order of |
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01:04 | 28 somewhere questions we'll have like 45 so more time to do these two |
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01:10 | it. So um kind of meant a no summary of the unit |
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01:17 | Okay um The uh there won't be five on there because you won't get |
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01:23 | until next week. We may start little bit of it on Wednesday, |
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01:27 | won't finish it until next the following . Um So I'm not gonna put |
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01:32 | on you know one but I may add some you know questions anyway. |
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01:38 | questions uh Chuck that relate to Chapter Material. I can I can put |
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01:43 | up next week. So um Let's . So smart work. No no |
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01:49 | do last week but this week. so to on Sunday three and 4 |
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01:54 | then two days later on Tuesday. um I just did it that way |
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02:00 | case you know you went through the and you had a question before the |
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02:04 | you could you could ask it. So um that's why chapter five is |
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02:09 | gonna not on a sunday. So due date give you extra time. |
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02:15 | See today so today I'm gonna finish three in the flip class. I |
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02:18 | my when I flipped in front of class it kind of makes people nervous |
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02:24 | something. It's like it doesn't count anything extra. Just just another class |
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02:28 | it's different that hopefully you're prepared for on your own. I'll certainly cover |
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02:32 | stuff but they're just aside from normal there's more questions clicker questions but to |
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02:39 | everybody's anxiety. If you have any clearer question you after today is two |
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02:44 | for this right or wrong. But do make a note of the |
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02:49 | you're getting wrong. Okay because you will want to rush up on that |
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02:55 | or ask questions about it. So do make a note of which |
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02:59 | ? You know if you get Alright then good for you but if |
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03:02 | get the market getting getting in one more of these wrong. Make a |
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03:06 | right? These all these um These posted after class. All all the |
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03:12 | clippers Tides, we do in this are always posted in lecture notes. |
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03:17 | a folder in there called it was extra. And or question question to |
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03:23 | something like that. So they're they're posted. So you you can look |
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03:26 | the clipper questions as you can for DLC today. Okay, so |
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03:32 | just make a note if you're you know, questions. Make a |
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03:36 | . Okay, so you can ask it later. The uh, |
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03:41 | So Mm Hmm. We need to is to finish up the last page |
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03:47 | Chapter three before we start. So 3 um we have now two |
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03:54 | which I'm not gonna say a whole about. Okay, so remember chapter |
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03:59 | . Is that the the continuation of the pro patriotic cell is and the |
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04:06 | and structures and so forth. so the last two things that cover |
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04:11 | are nano tubules very brief. And uh flag yellow. Some motion by |
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04:16 | yellow flag ela, pleural fluid Um singular. So now tubules um |
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04:25 | basically cytoplasmic extensions. You can see connections here between cells, basically cytoplasm |
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04:35 | between cells. Um, these extensions course, can lead to material being |
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04:42 | between cells. Okay. Uh there's lot really not known about this in |
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04:49 | of how they form. I just an article walking over here about |
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04:55 | that there is some dispute there. evidence showing that these might just simply |
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05:00 | a product of cell death in in bacterial species. So I'm not gonna |
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05:05 | things. We're just gonna go with your book is saying on this. |
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05:09 | just saying. There's a lot, not knowing about this and what function |
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05:13 | is. So all I can say this point and I'm going to say |
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05:16 | settle fascinating cytoplasmic extensions, connecting cells seen that can be of the same |
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05:22 | or different species. Um If you these kind of connections and anything in |
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05:26 | cytoplasm you know Saudi molecules, proteins be transferred between the two cells. |
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05:32 | . But again having what I just that were coming to class. Uh |
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05:36 | certainly some question about um how prevalent is when these forms etcetera. Okay |
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05:44 | that's already won a statement at this . Um You made those who are |
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05:49 | with congregation and they think okay well may be another way to congregate. |
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05:54 | . Okay. I don't I don't evidence is pointing that way. Congregation |
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05:57 | a very specific process. Gene control . Okay. I'm not sure how |
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06:02 | of this phenomenon is even controlled if is just a random thing or |
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06:07 | Okay so um I don't want to that much time on it more than |
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06:12 | . Okay. Um Do you want spend a little more time on |
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06:17 | So it's a gel um similar for plural. So that picture here is |
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06:23 | to show the bacterial types of the . Okay. Trouble things. Um |
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06:31 | bacteria. The motor. Our rod . Right. There's Maybe one or |
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06:37 | species that are cock sided Morimoto overwhelmingly remote style. Our rocket ship general |
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06:45 | are not moti Okay. Um and other broad shape types that are aren't |
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06:51 | as well but we're just gonna stick multiple ones, multiple biopsy jlo or |
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06:56 | for different arrangements of the project as see there. So even have 11 |
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07:03 | flagellum at one end of the you can have a group of them |
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07:07 | one end, you can have all the cell periphery. Um Or like |
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07:13 | guy right here, one on either . Okay. And that's probably not |
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07:18 | most common type of C. I'd probably this and this is probably the |
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07:24 | common. Um but uh I don't this but the one with one on |
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07:30 | and it's one or the other is functioning at a time. Not both |
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07:34 | , they both were functioning together. guess it would not be going anywhere |
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07:37 | a circle or something. So either or the other is functioning to make |
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07:41 | . Okay. Um the so gentlemen is made of uh units called gel |
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07:49 | like the pilot was made of pilon . This is made of Magellan |
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07:54 | The motion is quite different from what see in eukaryotic cells. Think of |
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07:59 | is um the movement is like a like action of the. Okay, |
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08:06 | motion is like a propeller. so very different. Uh and the |
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08:10 | is controlled by, what direction is moving clockwise or counterclockwise. Okay, |
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08:17 | two very different types of motion. The H. Antigen because he was |
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08:23 | about the number four with the gram OH. P. S. |
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08:28 | Um the H engine refers to the Ellen protein. So um uh the |
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08:36 | itself can be made that can have amino acid sequences in that fridge element |
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08:40 | makes it respond differently in terms of response. Okay so you can have |
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08:47 | . Both pendulum will induce an immune depending on their particular form amino acid |
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08:54 | that can change uh the evening So in other words as I talked |
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09:00 | before the O engine it's a way identify oh uh pathogen types. So |
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09:08 | has all been worked out the old A changing typically was was uh used |
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09:16 | identify different types of gram negatives like the E. Coli and salmonella |
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09:20 | There's hundreds of different e. Coli in terms of H. And |
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09:25 | Engines and it's all about being able identify medically important types and so we |
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09:30 | do this with and you see the 0157. Right? The E. |
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09:35 | we talked about before the old refers the O engine. There's an also |
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09:39 | H. Number as well referring to type of flu yellow. So many |
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09:43 | the important types we can I. . These using basically antibody engine type |
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09:49 | . Okay now the structure itself this showing a gram negative. I say |
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09:56 | because you see two outer and inner . Right? And so uh in |
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10:02 | this you'd have the uh what's called the basal body that anchors it in |
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10:08 | membrane. And the hook is where of it rotates gives us that rotary |
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10:13 | . It is of course energy It's not uncommon for a proton |
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10:19 | We talked about that before that, can supply the energy for this. |
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10:25 | The motion of this kind of pendulum all about um how it rotates, |
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10:32 | direction it rotates clockwise or counterclockwise. so that movement is determined by |
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10:40 | clockwise or chronic clockwise. It's determined the presence of chemicals. Right? |
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10:47 | doesn't have receptors for those chemicals. , so on the on one pole |
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10:53 | , nobody receptors there. And it respond to certain chemicals. Typically things |
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10:59 | that are called attractants, right? these are typically nutrients, type |
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11:04 | right? Maybe carbohydrates, maybe amino , you know, nutrients that can |
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11:08 | to grow. So these are kind things that serve as attractiveness. |
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11:12 | And we'll draw the sell the sell drawn towards them, right, Because |
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11:16 | have the receptor for it. and so the presence of these attractants |
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11:23 | the amounts that are present will influence type of rotation. Okay, so |
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11:31 | can have what are called runs counter rotations. So that kind of puts |
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11:36 | fort Gillem in if it's a group them all kind of in unison. |
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11:40 | , moving to sell forward or it be a tumble, if it's a |
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11:46 | rotation, that kind of unravels the ella. Okay, and in that |
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11:52 | this is kind of really the cell kind of just spinning in place, |
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11:58 | ? Not really going anywhere. Kind just doing this in place, I'm |
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12:03 | and so it's about the, adjusting frequency of counterclockwise and clockwise rotations. |
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12:10 | those proportions determine what kind of a it make a purposeful movement towards something |
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12:16 | is it just kind of randomly Okay, so what we're talking about |
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12:22 | is doing so, my rambling, random meandering is what you might call |
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12:29 | , right? A random walk which a higher tumbling frequency, a greater |
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12:36 | of clockwise rotations. Okay, and you see that the points here. |
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12:43 | , here, here, here these all tumbled. Okay, so you |
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12:49 | those mixed in with runs, The straight line movements. Okay, |
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12:55 | see lots of tumbles, all these here. Okay, mixed in again |
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13:03 | the front. Okay, and so , what proportion of those are we |
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13:08 | ? Okay, and so we have higher tumbling frequency than it really is |
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13:12 | of just randomly moving around, what's going on there? Well, there's |
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13:18 | very low level of attracting present, or no attractive is present in that |
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13:24 | . Okay, so think of it the cells attempt by going in different |
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13:30 | , right, so it's going uh it goes like in this direction, |
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13:35 | it goes that way, then it this way, right then that way |
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13:41 | goes down that way. So it's think of it as if it's doing |
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13:45 | randomly answering. Maybe the chances are it will run into something. |
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13:51 | Well we'll encounter attractant okay. And can influence the change in the proportions |
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13:59 | runs to tumbles tumbles two runs or look at it. So maybe we |
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14:04 | to something from here to hear. so this random walk that they call |
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14:10 | the left in the presence of attractive may turn that into a made bias |
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14:17 | right. Created bias. So it is going towards this is the kind |
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14:22 | terminology used by your book. But the point is as encountering attractants |
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14:29 | then increase the number of top counterclockwise , making more runs. And of |
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14:37 | attracted you see is um low lower to higher level is you can tell |
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14:46 | by the the color is more faded here. Okay. But it gets |
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14:54 | darker as we go that way. that's a gradient of attractive low to |
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14:59 | levels. Okay. Um And so it doesn't see there's a little bit |
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15:05 | a ton of randomness here right there there and there. But then think |
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15:12 | the triangles here as attractive molecules. , we're getting more and more of |
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15:18 | here. Okay and then as these sites fill up right then we get |
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15:26 | and more runs more and more Right. And so going toward you |
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15:31 | where there's lots of the attractive presumably you have to then you can |
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15:36 | Okay and grow um So again it's about the proportions of these two types |
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15:43 | rotations. One causes tumbles and kind random Mannering one creates runs and runs |
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15:51 | a attractive right? Um It's all three suckers, it has to bind |
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15:59 | attractive. It may not have receptors all types but for some Okay and |
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16:03 | it encounters it then potentially you can get that kind of move um Any |
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16:10 | about that? Yeah if it knows Okay so it binds random walk happens |
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16:15 | doesn't attract and so why does it have some? Because it's about well |
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16:22 | tumbling where there's the proportion of tracking still not high, it's kind of |
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16:27 | in the lower end of the Right, So you're still gonna have |
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16:30 | of that? Right? Not gonna completely but but so that means that |
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16:36 | me that not all the receptor sites necessarily filled up. Right. So |
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16:39 | still influenced some tumbling to occur. and and as it then keeps going |
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16:46 | higher levels of attractive then it gets and less. Yeah. Right. |
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16:50 | . Right. It was all about how much attractive relate college too, |
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16:58 | much of the receptors are being And so there's something they're not all |
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17:02 | and that can translate into maybe more . Okay, right, Right, |
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17:11 | . Yeah, of course. we have one. So we had |
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17:16 | clicker question uh last last time. we've got all through all these points |
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17:22 | . So let's see um uh how do here. Mm hmm. Thank |
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17:32 | . So there's your answering here. we have this So this will close |
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17:37 | tractor three. So there's a a of stuff there in terms of, |
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17:43 | know, the the appropriate excel. think of it as And it's easy |
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17:49 | test yourself on this is just simply a blank sheet of paper draw rod |
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17:53 | if you liked oxide drop big Right then think from the outside going |
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17:59 | , what what are you encountering? ? The envelope? What might be |
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18:03 | the envelope? Right? Is the negative gram positive? It's something |
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18:07 | Alright then you go to the membrane go, okay, what's what goes |
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18:11 | there? Right, well, it's of fossil lipids transport processes. Right |
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18:15 | you go, okay, what's Alright then you go, okay, |
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18:19 | , nuclear Oid, right, bribed own salutes, et cetera. Um |
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18:26 | we're talking about party zone formation. ? So I'm thinking of different |
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18:31 | We talked about the components and different of inclusions. Right? So it's |
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18:36 | way to think about it uh, test to test your knowledge on that |
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18:43 | . Um let's see here. so, um so I recommend doing |
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18:54 | . Alright, test yourself that Mm hmm. Okay. Here we |
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19:21 | . 321. Uh Yeah that is . So Magnotta zone is not a |
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19:31 | a food store that's all about orienting cells in space. So these are |
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19:37 | bacteria that have these and then um towards magnetic north or south. |
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19:42 | So it's kind of a movement. think of making some connections between these |
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19:47 | . Right? So example is think all the different things involved in that |
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19:52 | of motion, right? So we talked about flick gel and that's pretty |
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19:56 | to remember that that's about motion. so too things like the twitching motility |
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20:01 | and the pill I for that involved that a gas vacuum, right? |
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20:07 | move a foot a synthetic type up down in the water column. |
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20:10 | That's kind of that's motion certainly. maquina zones type of motion, |
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20:16 | So there's a lot of these different we talked about can relate to that |
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20:22 | That particular feature of most. Um okay, so like I |
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20:29 | lots of uh components and structures to appropriate cell. But do take that |
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20:35 | I mentioned terms of testing yourself, ? They go, okay, here's |
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20:39 | here's a cell appropriate cell. What's I'm going outside the end? What |
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20:44 | I encountering? What's going on? . So any questions. Okay. |
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20:51 | , this is just what we're talking motion. Right? Yeah. So |
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20:58 | is any kind of emotion. And typically you have either photo |
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21:03 | chemo taxes. Um There are other . I can think about the top |
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21:09 | my head but it's it's motion taxes the motion part. So photo taxes |
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21:14 | towards light. Chemo taxes mostly due the attraction to chemicals. So |
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21:20 | I think I'll answer your question. ? Yes. Chemo tax. |
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21:25 | Yeah. Exactly. Yeah. So as we go into Chapter four |
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21:34 | 1. All right. So the here, there's a number one theme |
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21:42 | is um growing growing bacteria. so how many in here are biotech |
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21:55 | ? All right. You will be cells. You're in biotech? |
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21:59 | Um The biotech even in of course I know you do gross sales and |
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22:08 | . You need to do some genetic stuff. Um I worked in biotech |
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22:15 | 12 years before coming to U. H. And that's what I did |
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22:19 | self. Well, more than That sounds pretty basic. But it's |
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22:23 | boil down to that because so when look at this overall, just kind |
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22:29 | thinking, okay, what do we do I care about that? You |
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22:33 | actually have these growth requirements. You them together to make the media and |
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22:37 | blah blah. Right. Um You grow them of course for different |
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22:43 | Right. So in any basic research um certainly you're studying different processes. |
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22:53 | look at proteins between sequencing and he's often not always but often involved growing |
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23:00 | cells. Okay. Or you can cells and tissue culture? Right. |
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23:05 | human cells can be grown that Other mammalian cells. Okay, |
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23:09 | basic thing about gross requirements. Of there is no different to grow skin |
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23:15 | and culture than to grow bacterial All of you have to be a |
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23:19 | they're a little more finicky more affinity grow um tissue culture cells because of |
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23:27 | way they are. Okay, a more complicated but still require the basics |
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23:31 | terms of growth. Okay. Um . And so again, growing them |
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23:37 | for different purposes, whether it's sequencing want I say DNA sequences, you |
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23:41 | proteins and biotech. It's often that grow themselves up to isolate through the |
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23:46 | of commercial value or or the cell , Maybe the commercial value. |
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23:52 | and when you grow things in lab quantities are probably fine. Okay. |
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24:01 | for commercial aspect that that will not you anywhere. Okay, now we're |
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24:06 | about thousands of gallons or liters. and um that's a whole lot of |
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24:13 | unto itself. So we're gonna start and keep it simple but we'll talk |
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24:17 | bioreactors. I have active growth here a little bit but it all starts |
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24:23 | , you know, what is What is the what's the requirement, |
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24:27 | requirements, putting those things together. What type of bacteria am I working |
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24:33 | ? Right. They're gonna be considerations Right, carbon source electron source blah |
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24:40 | . Okay. Um how do I to grow on solid or liquid |
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24:44 | Okay there's different roles for that. what kind of media complex defined? |
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24:49 | the Richmond blah blah. Okay so are all these decisions are all based |
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24:55 | what am I trying to do? if you're simply just trying to get |
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24:59 | ton of cells in a short period time then you need to know what |
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25:05 | what what what was it like to ? What are the optimal conditions? |
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25:10 | so maybe some before you go full grow something in 1000 liters. Okay |
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25:16 | requires some you know this this size on the bench top fiddling around with |
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25:22 | nutrients and whatnot. Which I'm getting curves and let's talk about growth curves |
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25:26 | the end. Okay so anyway so lot of this is uh you know |
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25:31 | of you will go on to make research lab, biotech lab and these |
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25:36 | the kinds of things you'll be Okay. Um Okay so there's one |
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25:43 | over, I should have told us time. Not a big deal but |
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25:46 | it's kind of the sequence. So dynamics of bacterial growth. That's kind |
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25:51 | the growth calculation stuff. We're not do that today. We're gonna save |
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25:55 | for next time. Okay I'm gonna through the meat of this stuff. |
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25:59 | is everything above above this. Okay it's gonna be a mixture of questions |
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26:06 | then I'll ask you something up. Not everything is a clicker question. |
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26:11 | you'll see questions written on the slide there's no A. B. |
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26:14 | D. E. It's just kind meant to give me the answer. |
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26:18 | me let me know what you Okay so let's start some kind of |
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26:23 | . Just kind of mentioned these points nutritional requirements. Physical chemical factors uh |
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26:32 | so physical factors like temperature, other like the oxygen concentrations may be |
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26:39 | Chemical factors certainly nutrient requirements. Growth formulation, putting those into a mixing |
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26:45 | together liquid or solid form. So growth media type complex are defined also |
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26:54 | have these categories selective differential enrichment Um Media formerly with solid and then |
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27:04 | I said a second ago we'll talk this um Next time. Cell numbers |
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27:10 | rate and those growth rate calculations aren't that's super complicated but we'll we'll do |
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27:16 | of those uh uh next time. so let's start here with this |
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27:22 | Okay so um for most bacteria increasing amount of this nutrient to a growth |
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27:31 | will typically yield a significant significant increase cell yield. Okay, which nutrients |
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27:38 | this be? Okay So kind of . No fast forward but then speeding |
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27:47 | up a little bit. You got time around? Mm hmm. Yeah |
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27:56 | question sure. Yeah. Just ask questions right as we go through |
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28:02 | Absolutely. Okay. Stop it at seconds. Didn't catch up. |
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28:14 | Mhm. Alright. I think 120 our headcount. Yes. Can |
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28:20 | 21 soup. Okay. Yeah, definitely carbon. Okay, so um |
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28:32 | , if you don't get these things , whatever all these side is gonna |
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28:36 | available to you afterwards, with the circled. Okay. Uh did you |
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28:41 | ? What was the question? I I have 80. It's kind |
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28:48 | nowhere when it comes to throwing some culture. So why wouldn't carbon |
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28:56 | The answer? Yeah. Because nitrogen X is actually number two. |
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29:04 | , carbon first election you you will out of um uh you can keep |
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29:10 | a culture of carbon carbon source and , right? Keep selling you going |
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29:14 | and up and up and up and can of course you'll you'll have in |
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29:19 | right you're C H. O. . P S. Right? Carbon |
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29:22 | oxygen, nitrogen phosphorus, sulfur those , right? Because those make up |
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29:26 | major biomolecules, right? You take , proteins, lipids, carbs, |
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29:32 | ? So, um but when you a media with those C H. |
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29:40 | . O. P. S, ? And you keep adding more carbon |
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29:45 | get more cells. You can keep for a long time before you run |
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29:48 | of nitrogen. Okay? So you at the carbon nitrogen ratios in my |
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29:53 | that's really what it's based on. why you need more carbon and nitrogen |
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29:56 | certainly, you know when you do these aren't things that you know when |
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30:01 | with this amount of stuff in the you don't really encounter these issues is |
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30:04 | you're trying to really get lots of for commercial purposes that that can be |
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30:09 | issue. And so you do find you do have to actually add nitrogen |
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30:12 | it's not until the cells are really very high density, correct. Um |
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30:18 | so we see the the six atoms . So you don't you don't really |
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30:26 | oh or h okay uh as part the you know additions because h is |
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30:35 | know in the hydrocarbon molecule right? always a joy around right as |
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30:38 | Oh typically um so what we do you know, nitrogen source of phosphorus |
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30:45 | sulfur source sulfur is a in one the amino acids and that's needed. |
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30:51 | macro micronutrients. So um macro or those things you see there. |
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30:58 | Um I typically differentiate in terms of add these in grams per leader. |
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31:05 | are typically a micrograms per liter Um so things like iron um uh |
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31:15 | elements is often called iron, nickel, tungsten. Um the growth |
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31:26 | , those are things you typically add you don't always know what's in there |
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31:33 | helps it grow. So like blood often used as an addition to grow |
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31:38 | . Um You just don't grow better you add these things because there's lots |
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31:41 | stuff in blood that they may be as a nutrient. So sometimes you |
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31:45 | go to the pains and figure out that is. You just know you |
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31:48 | that blood because you don't know they grow better um things like |
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31:52 | Um uh these can also be growth as well. So that's kind of |
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31:57 | that category is are typically nutrients that cell can't make themselves, You have |
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32:03 | add them. It's very often the . Okay now with sexual nutrients you |
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32:07 | essentially carbons and century because cell doesn't make its own carbon so you have |
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32:10 | supply it. So essential ingredients must supplied, right? Because the cell |
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32:15 | synthesize it. Right? Um Now and so this right here is sort |
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32:22 | the carbon skeleton. Right? Carbon life forms. Right? So um |
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32:28 | was a skeleton for all of your biomolecules and you add different atoms to |
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32:34 | . Uh nitrogen and phosphorus to make or to make it clear gasses, |
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32:40 | have you? Okay, but that skeletons the base, Right? That's |
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32:44 | I have so much influence and how material you get biomass you called |
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32:50 | Um many questions about that Yeah, I guess the I'll give you this |
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32:59 | . Car the carbon is the best , but all the other answers are |
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33:03 | kind of work as well, but as good as carbon. Right? |
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33:07 | . If you want to do if you had e coli here growing |
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33:12 | nutrient broth. Very basic. And it stopped growing because it just |
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33:19 | out of stuff to keep growing to itself. If you advise just plopped |
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33:22 | a couple of tablespoons of table Boom, they'll take off again, |
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33:28 | doubt because they wouldn't have exhausted all other all of these things. |
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33:36 | by that time, but you get carbon. There you go. |
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33:40 | so it's always number one not to higher, I'm sorry, hierarchy. |
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33:49 | uh I guess yeah, certainly carbon one not coming to Phosphorus, probably |
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34:00 | three, phosphorus 4. But I focus so much on sulfur. And |
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34:06 | from growing cells purposes, there's always on the carbon, your candidate |
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34:13 | Okay, next interesting next question. I'm gonna start the timer right |
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34:19 | So you got to read, read . Okay, so um agent deployment |
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34:25 | the chemically the chemical composition of various media Abc D. Which one is |
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34:33 | defined, I. E. Synthetic . Minimal medium is also called that |
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34:40 | suitable for the growth of a hetero pro carrier. So the operative terms |
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34:47 | basically uppercase and bold. Those are two. Too many things to no |
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34:53 | answer the questions. Okay um so what the head of growth is and |
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35:00 | chemically defined media is. Okay, you got the five recipes. Their |
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35:09 | . Take a look and let me that. Okay. Yes. Pay |
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35:36 | to this. Ah Next question Okay. Mhm. Alright, I'm |
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36:14 | put the timer on 30 seconds. . Three. Okay. Here we |
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36:48 | to 10. Let's see. So Let's uh six. Alright. Which |
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37:04 | ? We'll start with the first Which which is the chemically defined |
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37:09 | Mhm. Just anybody just give me choice but Yeah, correct. |
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37:19 | So uh so you see pep tone uh we have Pepitone, uh even |
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37:26 | yeast extract? Uh um pep Uh Right, so any medium that |
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37:37 | one or more of these in them a complex medium. Okay, so |
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37:47 | so one way to think of it if I give you a I think |
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37:52 | of these buttons probably pulls down the chart. They'll give you periodic chart |
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37:56 | a calculator. You could tell me are the grounds for most of all |
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38:00 | atoms in the media. Okay. can do that, you can do |
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38:04 | with D. Um E. Right? Because we you can write |
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38:09 | the chemical formula for glucose C six . 12 06. Right? And |
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38:13 | for the other things. And so can have a calculator periodic chart and |
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38:18 | it out. Right. Um The of these complex nutrients, you can't |
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38:25 | that, right? You know? Pepitone is Pepitone yeast extracts, says |
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38:32 | itself. It's yeast. Right? been treated to take all the stuff |
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38:38 | in yeast, right in its Okay, so what's that gonna |
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38:43 | Well, that's gonna be carbohydrates is to be proteins, nucleic acids, |
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38:50 | acids, right. Whatever is in yeast cell. Right? Is what's |
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38:53 | added to the media. Okay. just don't know what are the exact |
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38:57 | and the types of things in Right. Pep tone is basically |
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39:05 | Okay. It comes from a right? You go to a lot |
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39:09 | these ingredients whether it's beef extract or tone, a number of other |
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39:15 | You go to a slaughterhouse, you up the stuff that's on the |
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39:17 | Okay. Gross. I know. . But uh you boil it or |
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39:22 | enzyme treated what have you? All . And so you're basically getting all |
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39:26 | stuff that's in there. Right and meet and that's gonna be lots of |
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39:32 | . Right? It's gonna be me acids and and it's gonna be certainly |
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39:39 | right? C. H. N. P. S throughout in |
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39:44 | forms. Okay. But you just know where the exact importance of |
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39:49 | There's no chemical formula for Pepitone for . Right? So that tells you |
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39:52 | a complex nutrient and one of these the media makes the complex media even |
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39:57 | it's all if that's all it I can be right or even if |
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40:05 | looks like oh that's completely defined. right. The only added one doesn't |
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40:11 | just by adding that one makes the meeting. Okay and um that's those |
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40:19 | definitions now then the next part right so we know the chemically defined |
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40:25 | A. And D. And Okay. Sorry. Uh Okay. |
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40:36 | Yeah A. And A. And . Okay A. And D. |
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40:39 | it? So then which of those a hetero on hetero? Yeah. |
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40:46 | . Okay. Okay. So so it becomes the hetero trovato pro |
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40:53 | Right? So ahead of Trophy uses Hera tropes used only complex organic carbon |
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41:05 | . Right? So I used to and even textbooks still call it this |
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41:11 | inorganic carbon versus organic carbon. C. 0. 2 technically is |
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41:17 | carbon. All right. I got upside the head once by accounting professor |
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41:22 | 20 years ago for saying you have use inorganic carbon. It's not |
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41:28 | But How I frame it is CO2 the most the simplest Organic former |
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41:35 | Right? So it's c. 2 which chemically looks like that. |
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41:42 | . The super stable. You're not break apart those oxygen's right. Life |
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41:48 | do that, right? What life is takes the entire model kill and |
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41:54 | it as a building block to make complex organic molecules. That's what you |
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41:59 | that's what Auto Trophy is all about C. 02. Okay, taking |
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42:05 | and making complex molecules out of Okay? Which of course takes lots |
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42:11 | energy. Okay so fixing SOO two not a way for an organism to |
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42:15 | energy. It actually requires a lot energy to take SEO too. |
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42:22 | So Herod And you you can't do . You never forget but you're you |
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42:27 | ahead of you don't fix the Right? You eat these more complex |
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42:32 | like fructose, glucose, right? . More complex organic monitors. Okay |
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42:41 | header proof waterproof thing. Okay um the hetero trophy then we left with |
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42:48 | . And D. As our to defined types. Then the metro is |
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42:51 | to use this right? A not that's an autograph. So this tells |
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42:57 | that. Right? So we can can use you could bubble in |
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43:02 | 0. 2. That's one way another way is to add these carbonates |
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43:07 | you see here. Right? And solution that can be converted to |
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43:11 | 02 that can be fixed. So um anyway so this is basically |
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43:18 | hetero autotrophs difference here. Okay. the complex versus defining. So then |
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43:25 | look at um yeah go ahead. if the option is just carbon |
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43:36 | what If glucose were replaced with 2? Well it's there anyway. |
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43:45 | . It's in the it's in the . Okay so we're not you a |
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43:55 | you wouldn't if you're trying to get of trump you wouldn't put that in |
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43:59 | . Right. And if so to the ceo to any environment if you're |
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44:03 | tropic growing and it's not going to it. Right? But you wouldn't |
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44:05 | to carbonate to the medium. If you're trying to go ahead and |
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44:09 | I mean it would probably do anything it other than use it. You |
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44:12 | use it as a there's a non . 02 pipe plane. Okay. |
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44:18 | Yeah I'm trying to grasp the joseph kind of like in the same, |
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44:27 | . What are you you're ahead of . Do you fix the 02? |
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44:32 | do, wow. You're I'm gonna the paper on you. So it's |
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44:41 | completely different things. It just it be more different than night and day |
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44:45 | that point it's just it's just Yeah. Are you an autotrophs? |
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44:51 | you ahead of trophy? That's really it is. So it just comes |
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44:54 | to common sense. Well I don't about that but now trust me I've |
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45:02 | teaching us for so long and It's I guess it's not an easy I |
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45:07 | from my perspective it's seen as very know you should know that. But |
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45:11 | guess it can be a struggle and seen it it is a struggle for |
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45:14 | to wrap their head around it. . But um that's what I |
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45:19 | Do you fix co two can you no? Okay. You don't have |
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45:25 | don't have you don't have radios by wake our box lights in your |
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45:30 | You couldn't even try it if you to. That's the ends on |
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45:33 | okay so this won't be the last we talked about how to profile |
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45:38 | E. Okay so all right so the diagram that will read from top |
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45:46 | bottom. Right. All organisms then down. Okay. So, we |
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45:51 | of course classify organisms through metabolically classify if you will, uh by energy |
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45:59 | and carbon source electron source. And so we have a through |
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46:06 | Here A B C D E G. Are your choices? |
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46:10 | So, which box represents little Okay, let's go over the |
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46:22 | And so as you'll see, or you maybe know already that this |
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46:30 | you know, it can be the metabolic type. Can have multiple |
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46:36 | Mean the same thing. Okay, the trophy is one of those. |
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46:41 | , so his head or a You know, So, a lot |
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46:44 | these names can refer to the same of metabolism. Okay, So, |
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47:10 | can get it. I understand some these basics. Now, when we |
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47:13 | to chapter two it's heavy metabolism. , you will be ahead of the |
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47:19 | once we get there. Okay. on, mm hmm. Alright. |
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47:57 | seconds. Let it go. One . Okay. All right. |
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48:05 | Who answered? Uh D. Two . She's all right. She's pointing |
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48:14 | you and your D. So why you pick D. So, so |
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48:29 | what's a little trough in more basic is a little trip like you Or |
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48:37 | it like a more like a Which yeah. Didn't hear what you |
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48:50 | . Yeah, because why? the reason you picked it because it's |
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48:57 | here on this right here. Right that, right so little troves are |
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49:06 | . Okay there are different types of . They're not like a plant. |
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49:09 | a plant. Only in since they're a trophy. Okay. Pick SEo |
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49:13 | . But look at trump is one those that yes um is a chemo |
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49:19 | . Right? Chemo trough. So was chemical non light non non light |
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49:27 | occupations To get energy to fix the . Okay um Before I go |
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49:36 | What is which box is you or you on this um spectrum? |
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49:49 | Yes sir it quicker. Mhm. the. Yeah. Okay so we're |
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50:15 | which box are you? Yeah. . Yeah. Okay. Okay. |
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50:53 | . Alright. two seconds 1. . Yes. A. You are |
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51:03 | . Okay so they're getting from top bottom. Uh So the first category |
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51:09 | chemo photo right to the source. um And so the thing about uh |
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51:19 | metabolism involving C. Co. Two , right? You're taking auto programs |
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51:28 | those units and building up into a organic model of glucose and etcetera. |
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51:34 | so that takes lots of energy. so the difference between the photo chemo |
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51:40 | , Photo auto growth is what is energy source to supply that to be |
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51:46 | to fix the 02. Okay. takes lots of 80 PS. And |
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51:50 | . A. D. P. . 2 to 6 C. |
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51:52 | So the source of those two things come from light converting light energy to |
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51:57 | energy, which is what photo trips or from non light chemical oxidation is |
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52:03 | like Oxidizing H two, ammonium uh two S. Okay. Iron |
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52:16 | Okay. Um and other things. , this is what a little |
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52:22 | Or chemo autotrophs or or chemo Okay. Um so I said you |
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52:28 | these names can be combined and refer the same type of metabolism. Let |
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52:34 | show you what I'm talking about Okay, so there's there's various |
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52:38 | right? Um and so again repeated . Okay, so categorized based on |
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52:44 | of carbon energy, electrons. So combine these terms, right? You |
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52:49 | have a a chemo lift. A , a chemo organa trophy. |
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52:54 | Um the terms you see there and are some of these this and this |
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53:01 | synonymous. Okay, this and this synonymous. Okay, the photo editor |
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53:09 | . Alright, okay. For the tropes use carbon dioxide that just yes |
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53:16 | no anybody say no. They say . Right. That's right. That's |
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53:26 | right. That's the operative term. ? So you see because everybody |
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53:32 | the your Mhm. You see a plant. Alright. Of course. |
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53:39 | this is the operative term. It's a you have to have you |
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53:47 | use light but it still needs more complex organic forms. In fact |
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53:55 | fix you too. Okay, so let's see, I think we're gonna |
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54:04 | on this theme a little bit. let's look at this next question. |
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54:09 | . Um So what kind of growth is this? Okay, so everything |
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54:14 | that box is in this growth Everything in the box. So the |
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54:22 | we had a few slides ago, should be able to get this. |
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54:32 | hmm. Mhm. Yeah, that , I Yeah, well, that's |
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54:57 | fine. That showed well, will available. I'll have that like the |
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55:03 | that he asked the question on like A. B. C. |
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55:06 | E. F. G. that's how it'll be. It'll be |
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55:09 | be I'll have it available. It'll be it'll be in this |
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55:13 | Yeah, that is a complete Yeah. Yeah. I have some |
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55:18 | the boxes covered up, but I'll them. So you see the whole |
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55:22 | , but but it's not more to than that. Yeah. Okay. |
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55:28 | think uh let's do be at the here. 765-4321. Mhm. Uh |
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55:44 | , complex we need. Alright. talked about these complex nutrients. So |
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55:48 | has Pepitone and beef extract. Um and so the so I have |
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55:57 | complex nutrients. Well, it makes medium easy. Right? You're your |
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56:05 | biologists, basically what we call, , They've developed a media called |
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56:12 | Which is basically like three complex if I remember. Right? |
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56:16 | Um And it means it's very so you have to weigh out, |
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56:23 | know, they don't have to weigh this and this and this and this |
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56:27 | this Alright. As you do for defined media, Alright, just add |
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56:30 | that boat. We're done okay. you know, you're supplying again, |
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56:36 | alone has C H O M P in it. Okay, So does |
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56:41 | we also know the exact proportions were owners. So we know it's in |
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56:45 | and the cells will grow okay. realistically, um if you are |
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56:51 | my familiarity with me is is always combination of the two. So, |
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56:55 | like this, because you want to really, these are actually not |
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57:00 | These aren't great carbon sources to get yields. So you typically want to |
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57:04 | something like this to add to add it. So you can get your |
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57:09 | numbers up high. Okay. But know, the pep 20 beef extract |
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57:15 | ? Because they're additional things like you , when the cells have to do |
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57:19 | for itself. Right? And by , I mean turn genes on to |
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57:23 | pathways onto blah, blah, Okay, that takes energy. It |
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57:27 | time. All right. So, you can give some of the stuff |
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57:30 | sells, you know, then they'll faster, Right? Because they have |
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57:33 | make that stuff for themselves and they quickly get up to yield and just |
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57:39 | this, manipulate your carbon source. , To get lots of cells. |
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57:45 | ? Could you give him some of stuff down there is complex insurance to |
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57:48 | them some, you know, a . So then we have to make |
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57:52 | the stuff themselves. But if you interested in finding out what are the |
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57:58 | nutrient requirements of these cells. Okay you take out this because you do |
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58:06 | to know what the exact amounts of in there because you're trying to figure |
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58:11 | . Okay, what if I manipulate or that? Will I get better |
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58:14 | or whatever. Right. So there certainly cases where you want to know |
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58:20 | in there be have control over all it. Okay. That's when you |
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58:24 | just have a defined minimum media. , so again, there's purposes for |
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58:31 | different things. Right. And so differential. Okay, differential. Simply |
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58:39 | medium. Usually a solid medium Okay. Where when you throw selves |
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58:47 | it bacteria, they will produce different reactions. Right. And that will |
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58:53 | you something about the physiology. Very common gram negatives. Just to |
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58:58 | media that has like different sugars on . Like say a medium with |
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59:04 | And you want to see does it glucose? Right, fermentation czar often |
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59:09 | acidic. So you can have some of ph indicator in there that when |
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59:13 | turns acidic it turns that color It's very common to look at |
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59:17 | Is that way on a plate? , if it's if it's positive, |
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59:22 | it will become acidic and will turn ph indicator. Read write those that |
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59:27 | ferment and won't become acidic? They on colored. Right? So you |
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59:30 | of see a difference. Right? it gives you a visual on a |
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59:34 | to see something. Tell you something its physiology, right? Um selective |
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59:41 | . Right? Is for example it be something like like this formula over |
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59:50 | . Okay. It could be All right. But then let's say |
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59:55 | add mhm bile sauce. Right. that can inhibit gram positive bacteria. |
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60:06 | , so you're actively adding a chemical inhibit certain types from growing do you |
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60:13 | gets elected? You can add an that will certainly be selective. Okay |
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60:19 | selective media is you're actively adding something it to prevent something else from |
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60:23 | Okay. And you can have You can have just selective you could |
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60:30 | just differential or you can have both . You can as a selective agent |
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60:36 | something out but then make it differential you're looking at some kind of biochemical |
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60:41 | that they have or don't have. , so you can combine both of |
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60:46 | . Um Any questions about that? a specific policy bios also won him |
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60:55 | gram positives. Yeah. That's Uh huh. Yeah. Yes. |
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61:06 | . You can have I'm saying in growth medium you can you can be |
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61:09 | selective and differential. It could be selective or it could be just |
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61:16 | That could be three. Could be the other both in the media. |
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61:24 | . Um Generally the p well in in a solid medium, like for |
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61:31 | test like that, we're looking for . The it won't be necessarily the |
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61:36 | inhibitory, it can be growth inhibitory liquid because the ph of the surrounding |
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61:40 | is getting Um there's of course affect and can cause growth to two slow |
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61:48 | . But generally that they didn't get a level high enough where before it |
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61:53 | to affect them. But the bottom is yes, if a culture is |
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61:57 | acidic, it can get to a where it can affect growth. |
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62:02 | Um there's ways you can get around in liquid culture if you need |
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62:07 | Okay, there's ways to kind of that you can control ph and the |
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62:12 | . You have everything under control. can control ph that way by neutralizing |
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62:15 | . We'll talk about that a little any other. Yeah. So for |
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62:21 | ph also. Mm hmm. I mean, yeah, certainly a |
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62:32 | now the only time you would you want to increase ph would be if |
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62:38 | were trying to grow an organism that that ph so those types of that |
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62:46 | cinephiles and what are called athlete files no basic or acidic ph But now |
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62:53 | gets into the realm more of enriched gets a hold that thought I'm gonna |
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62:58 | about that in the slider to there's a subtle difference between productive and |
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63:03 | media. Okay, so let me let's go to this question here. |
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63:09 | so this is actually a not a question but um what about term known |
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63:16 | be a Histamine oxide troll that grow that medium? Forget just take this |
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63:24 | of here. So we'll grow on medium here. So you gotta remember |
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63:29 | oxide trophy is. Right. So anybody care to guess would be able |
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63:36 | grow with 15 oxygen growing. Thank Wild guess. But anyway, |
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63:44 | Yes. I don't think it It's true. Okay. What's what's |
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63:51 | oxygen? What's the history of proof ? Right. It can't grow in |
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63:57 | . So not some trophy is deficient a metabolic pathway. Very common to |
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64:03 | a amino acid off control for some um Where you have to add that |
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64:08 | acid too along with the growth. if we have hicks history and oxygen |
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64:14 | that means it can't grow without the of history. Okay so knowing that |
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64:20 | be able to grow in this he says yes. Why? |
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64:31 | Yeah. Yeah. Those those complex have all that stuff in there. |
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64:35 | have to remember we have confused a extract or Pepitone is meat. |
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64:42 | Meat but that's really what it Um So what is what is what |
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64:48 | this poor example of? Okay what this? That's fat that's not uh |
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64:58 | you know it's gonna have everything in . We're just gonna have vitamins is |
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65:01 | have um proteins and all that Right. So so yeah. So |
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65:07 | it will have facilities in there. it shouldn't be able to grow on |
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65:11 | . If you took the complex nutrients , took a pepto and beef extract |
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65:16 | , then it wouldn't grow like unless have it. It has to be |
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65:21 | . Um Yeah, the Yeah. questions about that. Yeah. |
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65:31 | mm hmm. Also was the first of that uh It goes to |
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65:40 | Thanks to her question about ph so hold that thought we're almost there. |
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65:44 | . Um Okay, so, I'm gonna do some kind of beating a |
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65:48 | horse with this thing. So, just do let's just do um this |
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65:55 | . Okay, So which medium would chemo organic trophy? Go on. |
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66:00 | , you got three media types. , let me move that. |
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66:04 | B. And C. Okay. There's the L. B. I |
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66:10 | I alluded to earlier uh M. L. B. M. |
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66:13 | And that sulfur oxidizing media abc. remember chemo, chemo or gandolfini terms |
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66:23 | have multiple terms that refer to the same. All right. Um |
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66:43 | mm hmm. Okay. Timer's Okay. The truth. Mm |
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67:49 | So, the key to these these of questions is always focus on each |
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67:55 | the media types. There's one thing really ministry focus on and what's probably |
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68:02 | ? Mhm. Which one? Carbon source focus on the carbon what's |
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68:07 | carbon in the medium that will need to the right path track. Um |
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68:15 | so let's see. Mhm. So the majority answer. D Which |
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68:24 | correct? Okay so key morgana trolls I have actually have different then get |
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68:33 | you need I'm not gonna do those questions but but this this and this |
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68:40 | the same. Alright. See morgana hetero same thing. Right? You're |
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68:46 | both of those things. Okay. And so that would be um |
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68:54 | T. And B. Okay, autotrophs. We just talked about that |
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69:03 | second ago, right? They're both same as well. Okay, so |
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69:08 | gonna be medium C. Okay. C. C. Okay so |
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69:17 | carbon source. Right. C. . 2. Alright. These have |
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69:22 | this this to these can serve it's C. H. O. |
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69:27 | . P. S. When you complex nutrients, they have all of |
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69:30 | . Right? So they can be carbon source not great, but they |
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69:34 | serve as a carbon source. you can eat proteins, right? |
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69:39 | have to just eat sugars, you eat proteins. You need to click |
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69:43 | , You eat those things. Um , any questions about that? |
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69:51 | so focus on the carbon on these of things. Alright uh We talked |
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69:57 | this um you know, one thing will say is um with solid media |
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70:05 | wouldn't use solid media as a way get lots of cells to do stuff |
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70:10 | . Right? Because it's just not and it has a different purpose. |
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70:16 | solid media is some in some form fashion. Solid media plays a part |
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70:22 | pure culture for those that you can can cultivate the lab and isolate. |
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70:28 | your only hope to get pure culture a plate because you can, if |
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70:33 | have liquid right, you can have course have many different types of cells |
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70:37 | there, depending on the nature of sample. But you cannot you cannot |
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70:43 | can look under a microscope at that sample. You can maybe visually |
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70:48 | oh yeah, there's one swimming not ones that rod wants a |
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70:52 | Someone's this and with that. But all you can do. Unless you |
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70:56 | a pair of magic tweezers to pull out and then separate everybody and grow |
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71:01 | . You know, it's not gonna . Obviously you can't do that. |
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71:03 | you have to get this on the . Okay. And then they'll grow |
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71:10 | colonies and maybe you'll see different And then you can go, |
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71:14 | there's my different types. And I now take this guy and put them |
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71:17 | this plate. And this guy on plate and you get pure cultures |
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71:21 | You can only do that with solid . Okay. The utility of the |
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71:25 | of course, is to get lots cells. You can adjust your volumes |
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71:29 | from one mil 200,000 m if you to. All right. So it's |
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71:35 | about getting volume. Lots of cells liquid. Use liquid called the cultures |
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71:39 | to measure growth. Okay. Um , you know both of them have |
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71:45 | roles and depending on what you're trying do. Um Okay, so, |
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71:49 | to the that's what we got So, enrichment culture. Okay. |
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71:55 | um so this is a trick So, just uh so the hint |
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72:01 | is this here's the hint. um So the key is what's what's |
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72:08 | from that medium? Is there is something missing from that growth medium? |
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72:13 | what you know about um uh what needed in a growth medium? There's |
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72:21 | missing. Yeah, the action is . Okay. So what's going to |
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72:27 | on there? Well, this will , right. We're called nitrogen |
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72:34 | Okay. And two fixing. And doesn't write this down, but it's |
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72:41 | about let's talk about actually cycle later the semester. But they are a |
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72:45 | of bacteria. Very important. And very way to enrich. So |
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72:51 | versus selected million. You mentioned You mentioned ph Right. So, |
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72:58 | can use those parameters to enrich for types, Right? Because you could |
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73:03 | could use elevated temperatures to to enrich thermal files. Right? The elevated |
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73:10 | . So at that temperature you'll favor growth over others. Okay. Similarly |
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73:15 | ph right, you wanted to cultivating cinephile. Right? You can have |
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73:20 | medium at ph two or something and going to be enrich for those |
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73:25 | Okay, um or this meeting. ? We want to enrich for nitrogen |
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73:30 | types in the soil. Take out 19. That's what will allow their |
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73:35 | . So some of the difference between and selective selective media that to me |
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73:43 | you are adding a inhibitory chemical To stop a certain class of bacteria |
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73:50 | growing. Okay, uh enrichment. trying to favor the numbers on your |
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73:58 | or growth medium over others. Um in a handful of soil, |
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74:05 | you were trying if you just use medium like that, okay, you |
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74:13 | a ton of stuff growing on that , right? Because it's going to |
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74:18 | the needs of a lot of stuff that sort, Right? If you're |
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74:21 | to weed out, I want to a fixer in that group. We |
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74:25 | a hell of a time trying to that thing out of all the stuff |
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74:28 | going to play. So what do do? Well, you can enrich |
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74:33 | those numbers by growing on something like , then you'll get all the other |
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74:38 | out of there. Okay, um is no chemical you can add to |
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74:44 | all those without may be affecting your fixers from growing. Right? So |
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74:48 | so in Richmond selective. Right? in selective medium you are making that |
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74:53 | the sole purpose of I don't want negative growing on this or gram positive |
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74:58 | in this or I don't want So are sort of chemicals you can add |
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75:01 | inhibit those types, right? Some you might and antibiotic, right? |
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75:07 | in enrichment culture you're not really adding chemicals. You're you're setting the conditions |
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75:15 | favor the growth of a certain type that environment. Okay. Over others |
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75:21 | they may not they may not be to outgrow. There's other types. |
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75:26 | you just use a conventional type of , you have to get more enriched |
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75:30 | conditions for them. Okay. Does makes sense? Yeah. Um so |
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75:38 | so the part two is relatively We're just covering and those spores and |
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75:43 | in chapter four Part two. So have lots of time to finish up |
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75:47 | we got here, which is the curve growth curve and then some growth |
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75:52 | . So we'll have plenty of time time. So thanks folks and see |
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75:58 | on Wednesday. And this stuff will |
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