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And so instead of having the fragments where we stopped on the A's, we're going to have the fragments where we stop on the g's. And so, because we only have two CS, so this C doesn't count because it's part of the primer. So we have one C and we have a second C. So the complementary would have a g here and a g here. So we only form two fragments. And this lane would contain two bands because we only contain two fragments with different sizes. Then we repeat the process. | Sanger Sequencing of DNA .txt |
So we only form two fragments. And this lane would contain two bands because we only contain two fragments with different sizes. Then we repeat the process. Instead of using this one, we use DDCT. So C, that means we have to count up the GS here, not including this one, because it's part of the primer. So we have one g, we have two GS. | Sanger Sequencing of DNA .txt |
Instead of using this one, we use DDCT. So C, that means we have to count up the GS here, not including this one, because it's part of the primer. So we have one g, we have two GS. And that means we're going to have two complementary CS. So we're going to have two different fragments once again, one, two. And finally, if we use this one, we have to look for our adenine. | Sanger Sequencing of DNA .txt |
And that means we're going to have two complementary CS. So we're going to have two different fragments once again, one, two. And finally, if we use this one, we have to look for our adenine. So we have to look for the adenine. So we have one, two, three and four. We should have four fragments. | Sanger Sequencing of DNA .txt |
So we have to look for the adenine. So we have one, two, three and four. We should have four fragments. And that's exactly what we get in this particular case. So basically, in step three, we take step two and we repeat that same step three different times with the other three Ddntps. And in step four, once the four reactions are completed, we run gel electrophoresis. | Sanger Sequencing of DNA .txt |
And that's exactly what we get in this particular case. So basically, in step three, we take step two and we repeat that same step three different times with the other three Ddntps. And in step four, once the four reactions are completed, we run gel electrophoresis. Each reaction mixture is placed into a lane. So lane one, lane two, lane three, lane four. And the results are then transferred onto a polymer sheet. | Sanger Sequencing of DNA .txt |
Each reaction mixture is placed into a lane. So lane one, lane two, lane three, lane four. And the results are then transferred onto a polymer sheet. And then we use X ray order radiography to basically determine exactly where those radioactively labeled fragments actually were. And so this is the diagram that we get. Now, how can we use this to actually determine what the sequence of that initial DNA molecule is? | Sanger Sequencing of DNA .txt |
And then we use X ray order radiography to basically determine exactly where those radioactively labeled fragments actually were. And so this is the diagram that we get. Now, how can we use this to actually determine what the sequence of that initial DNA molecule is? Well, we know what the first three nucleotides are because that's the primer. So we have T, G and C. So the question is, what are these remaining nucleotides here? Well, let's try to use the following setup to base determine what the sequence is. | Sanger Sequencing of DNA .txt |
Well, we know what the first three nucleotides are because that's the primer. So we have T, G and C. So the question is, what are these remaining nucleotides here? Well, let's try to use the following setup to base determine what the sequence is. So we know that the farther down along our page, the the smaller our fragment is and the closer the nucleotide sequence is to the beginning. And the fragment all the way at the bottom basically describes this right over here. So that means the first nucleotide following the primer is the one lowest at the bottom. | Sanger Sequencing of DNA .txt |
So we know that the farther down along our page, the the smaller our fragment is and the closer the nucleotide sequence is to the beginning. And the fragment all the way at the bottom basically describes this right over here. So that means the first nucleotide following the primer is the one lowest at the bottom. So this and according to this graph, according to this setup, the lowest one at the bottom corresponds to a T. So this one is a T. The next one is we have an A. The next one is so let's make it a little bit bigger. We have a T. We have an A. | Sanger Sequencing of DNA .txt |
So this and according to this graph, according to this setup, the lowest one at the bottom corresponds to a T. So this one is a T. The next one is we have an A. The next one is so let's make it a little bit bigger. We have a T. We have an A. Then we have a T. Next we have, next we have a C. Next we have a T. Then we have two A's, one and two. | Sanger Sequencing of DNA .txt |
Then we have a T. Next we have, next we have a C. Next we have a T. Then we have two A's, one and two. | Sanger Sequencing of DNA .txt |