00:01
So this question says, do we need reverse transcriptase in pcr reaction? the answer is no, the pcr reaction itself does not need to have reverse transcriptase.
00:11
So let's introduce the pcr reaction itself.
00:15
So pcr stands for polymerase chain reaction.
00:19
So the idea is that you start out with a dna template.
00:29
So you have a double -stranded dna.
00:31
Now you want to amplify a certain region.
00:37
So to do that, you will actually need to have first of all, primers.
00:46
So the heat will be used to separate the dna strand.
00:50
So you can see you now have two single -stranded dna.
00:58
Then you will design primer that will be complementary to the dna template.
01:07
So you will have to know the sequence of the region that you want to amplify.
01:12
So you get two primers.
01:13
One is reverse primer.
01:15
The other is forward primers.
01:23
So the primer will be complementary to the region that you want to amplify.
01:28
And then you can see the arrow show you the direction of amplification.
01:31
So the primer will anneal to the template.
01:40
Now, after that, you'll actually have a new nucleotide dntps, and you also need tac polymerase.
01:54
Tac polymerase is a heat -resistant dna polymerase.
01:59
So the idea is that once the template, the dna polymerase, which is isolated from a thermophilic bacteria, will work to add the nucleotide to the end of the primer, the three -prime end of the primer.
02:24
So you can see that this is the primer, and the dna polymerase, tac polymerase is going to add the new nucleotide at the three -prime end of the primer and extend this way.
02:41
And this enzyme is tac polymerase.
02:47
So this is the key enzyme that actually adds the nucleotide to the three -prime end of existing primer.
02:55
So as you can see that no reverse transcriptase will be needed.
03:00
Instead, you need enzyme tac polymerase.
03:03
And so this is the same with the other strand.
03:06
Primer first, and then new nucleotide is being extended from five to three directions.
03:14
Again, the tac polymerase will do this job.
03:18
Now, so after you finish this round of replication, you'll end up having two double strand dna, one old, one new...