00:01
Explain the importance of rtpcr for amplifying a retrovirus, a viral gene.
00:09
So a retroviral gene, for example, hiv, have their genetic material stored as rna, but not dna.
00:57
So this means that since it's rna, the traditional or standard pcr can only amplify dna sequence.
01:28
Because dna polymerase, which is the enzyme that used in pcr, cannot use rna directly as a template.
01:47
So in this case, we must use rtpcr to actually basically amplify rna sequence, or to, i would say, to identify or isolate the rna.
02:04
So from here, for rna, you must use rtpcr.
02:09
So let's talk about why, how does rtpcr work out? so first of all, the retroviral rna is being extracted.
02:42
Now, then you would use the rtpcr kit to convert all retroviral rna into a complementary dna, or we call c dna, use the enzyme, a reverse transcriptase.
02:58
This usually is in the kit.
03:00
So this enzyme, which can convert rna into c dna or complementary dna.
03:52
So for example, you have rna, and then after you convert into c dna, it basically is, sorry, this isn't here.
04:46
So you can see that we produce a c dna according to rna by you.
04:53
Using reverse transcriptase.
04:57
So you can see that the cdna, the bottom strand, basically mimic the rna.
05:04
So this is the coding strand, which actually mimic the rna.
05:09
So basically, cdna mimic rna the sequence.
05:12
They are almost identical, except that you have t in dna versus you in rna.
05:18
And it's a double -strand structure.
05:20
So this is what we call reverse transcriptase...