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Kelowue frugnen o DNA_ The DNA hasdifferent restriction Sc;How MTAMY fragments would be generated CYou digest with only A"enzyme that recognizesHow many fragme...

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Kelowue frugnen o DNA_ The DNA hasdifferent restriction Sc;How MTAMY fragments would be generated CYou digest with only A"enzyme that recognizesHow many fragments would be generaled if you digest with Cn{Vme Lhal only recognizes B?Hou MlunI [rugments would be generated if you digest with Lwo enzymes pnC [nul recognizes tne and the other that recognizes B?How IAnY fragments would be generated hcre Wus muatOn the restriclion sile such that the restrietion 00zyme no longer recognizes that scqu

Kelow ue frugnen o DNA_ The DNA has different restriction Sc; How MTAMY fragments would be generated CYou digest with only A" enzyme that recognizes How many fragments would be generaled if you digest with Cn{Vme Lhal only recognizes B? Hou MlunI [rugments would be generated if you digest with Lwo enzymes pnC [nul recognizes tne and the other that recognizes B? How IAnY fragments would be generated hcre Wus muatOn the restriclion sile such that the restrietion 00zyme no longer recognizes that scquence for cleavuge? Using the box below rcprcicni agarose gel, dr# predicted restriction fragment pattems below' for digesls deseribed 4-4 Label the gel with and indicate which cnd the positive und negative electrode. De sure label your lancs. Ak muia Loading end



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When DNA is subjected to electrophoresis in an agarose gel, shorter molecules migrate faster than longer ones. Closed-circular DNAs of the same size but different linking number also can be separated on an agarose gel: topoisomers that are more supercoiled, and thus more condensed, migrate faster through the gel-from top to bottom in the two gels shown below. A dye, chloroquine, was added to these gels. Chloroquine intercalates between base pairs and stabilizes a more underwound DNA structure. When the dye binds to a relaxed closed-circular DNA, the DNA is underwound where the dye binds, and unbound regions take on positive supercoils to compensate. In the experiment shown here, topoisomerases were used to make preparations of the same DNA circle with different superhelical densities $(\sigma) .$ Completely relaxed DNA migrated to the position labeled $\mathrm{N}$ (for nicked), and highly supercoiled DNA (above the limit where individual topoisomers can be distinguished) to the position labeled X. (FIGURES CAN'T COPY) (a) In gel $A$, why does the $\sigma=0$ lane (i.e., DNA prepared so that $\sigma=0,$ on average) have multiple bands? (b) In gel $\mathrm{B}$, is the DNA from the $\sigma=0$ preparation positively or negatively supercoiled in the presence of the intercalating dye? (c) In both gels, the $\sigma=-0.115$ lane has two bands, one a highly supercoiled DNA and one relaxed. Propose a reason for the presence of relaxed DNA in these lanes (and others). (d) The native DNA (leftmost lane in each gel) is the same DNA circle isolated from bacterial cells and untreated. What is the approximate superhelical density of this native DNA?

So that Joe is going to be seen as follows where the number of killer bases, like 12 97 such are on the right on the different mediums. Like, indeed, we're gonna be in the middle here. Oh, so that we can see you. I wouldn't reply places they are as to be able to locate them.

Already. So here we're looking at, um, genetic analysis and in particular, the electrophoresis. So we have a, uh, piece of DNA, which pants sell sort of put off to the side and vertically. And three ability to sort of split it. We're using the restriction sites at with a two que be peace A for KB size piece killed this piece and a five. All right, CBI size piece. And so part A says this piece of DNA is cut just in this normal expected distribution that we are given in the problem. So what will the gel look like? So, for part A, we're gonna get lines at the length of these pieces. We would expect to see one at 21 at four. And one at five. Okay, So then part B says so. It's a mutation alters at site one. Okay. And site one was his first one. Now all of the banding look like Well, so there's a mutation there. Then it's not going to get cut there. Okay, so this first piece ends up being six k b in length, and then we start the second piece, which is five. So then we head onto part. See similar question. Let's say there's a mutation in both the first and the second one. So now we're not getting this DNA cut at all. The total length then is going to just be 11 and that that's it for that piece. Okay, so then we go into D. And this one says, if 1000 base pairs of DNA were inserted between the two restriction sites, So now we're adding 1000 to this one. What would the banding look like now? We're assuming that both of these cuts are going to occur still, So we still have our to K V and R five K be paid pieces that we need to add 1000 to the four key piece, but that's gonna make it five as well. So we're gonna put two bands at the five mark, And so what it would look like to the technician is just the two band showing up and then last but not least, we're adding 500 base pairs. We're not adding or subtracting. So 500 base pairs between the two restriction sites were deleted. Then what happens? So instead of adding 1000 were subtracting 500 s in this case, we're still gonna get R two and R five pieces, so we'll just go ahead and write those than the four were, you know, subtracting 500 from. So instead of being at four, it's gonna be at, like, 3.5, so be a little bit lower than the one in a

At the december might. Yeah. Athenian bromide. Yeah. Yeah. Yeah. Yeah. E. T. We are is sometimes I did to running before 82. Mhm. The running buffer during the separation of DNA. Okay. During the mhm. Mhm separation of DNA fragments by a gross gel. Mhm. Okay. By a gross yeah. Yeah by agro gel. Yeah electrophoresis. Yeah electro four aces. It is used to it is used because upon binding of molecules Okay because binding of molecules. Mhm. Because binding of molecules DNA. Thank you. DNA. And illumination with UV light source. Mhm. And yeah. Mhm illumination did. Mhm. You? Re light source. The DNA banding pattern can be visualized. The DNA. Yeah bending patron together can be Oh visualized visualized. Okay. Yeah visualized. Mhm. And this is the diagram of band appears on jail. Mhm beds still up there. Mhm. On Jason diagram and this is wealth DNA bench largest and smallest bands.

So in this podcast we're drawing out deal Drifter restriction fragments and the Crimson. So firstly, what we have is following. So here we have nine one, five, three. So for to six, um eight. Drive it Too long. Eso Now we have between nine and seven. Change the color hit. So between nine and seven. Here we have a between five and eight and we have B between one and four. We have see between five and two. We have t between nine and one. We have feet in between For Andre to we have a


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