I`m trying to understand the philosophy of primer-design for RT-PCR. Here I read some very useful posts (many thanks!), but still have few general questions:
1. template DNA means the antisense, non-coding strand, which is complementary to the sense mRNA transcript that is translated into protein;
2. non-template DNA means the sense, coding strand, which does not code RNA but is like mirror image of the mRNA transcript. And this is the published sequence in the databases. It gives antisense RNA transcript, which is not translated into protein;
3. if this is correct, complementary to the sense DNA will be the reverse, downstream, or anti-sense primer;
4. in parallel, complementary to the anti-sense DNA will be the forward, upstream, or sense primer.
Is that correct?
Edited by Nephrit, 14 January 2011 - 02:24 AM.