Determine specific primers and transgene copy number by PCR - (May/08/2008 )
I am trying to genotype my potential mouse founders. I understand i need to optimise PCR conditions, so first
1. I want to standardize the primer conditions, if i use any genomic DNA and do a PCR with my specific primers, I am not suppose to get any band ( as the genomic DNA is not ffrom my transgenic),
a) then what do i expect in results (on gel)
and how this will help in primer-standardization ?
2..I know one can detect the copy level also by PCR, how to standardize the PCR conditions to know that it is sensitive enough to detect the transgene at a single copy level?
3...Can anybody share a picture of genomic DNA ran on a gel, its a big DNA (compared to plasmid, which i have worked on), how does it look on gel..a smear..a clear band like plasmid DNA...
Any input is appreciated.
See the first result: