Hi there PCR friends,
I need help designing pcr primers! I HAVE NEVER designed any Pcr primers before.This is part of my MSc project
This is what I want to do:
1) Design HRM Primers for 2 virus (HSV 1/2)
2)The primers have to prime for both virus species.
3) I will differentiate between HSV1 and HSV2 by melting analysis.
4) I would like to include an internal control that would be primed by the same primers and this DNA can be synthesised.But it has to have a lower melt than HSV 1/2.
5) Include a human housekeeping gene (e.g. actb) to make sure that we have human cells in our sample. I would use HRM too here.
1) All of the above but using different primers for HSV1 and HSV2.
Where should I start?
I have been on the NCBI database and retrieved the viral sequences.Where to go now?
I would like to learn how to do it and just need pointers in the need direction!