Hi everyone
I apologize if this is a basic question but I am so confused.
I want to do PCR-sequencing for the human-gene which has some psuedogens with high sequence similarity (97%) with the original gene.and I have no choice except designing primers at the regions with 1 or 2 mismatches from the psedogenes...
so I'm not sure if the psuedogenes would amplify to some extent with the original fragment or not at the end
I noticed that the orientation of the original gene at GeneCards browser is:
minus strand (Start: 2,138,711 bp from pter End: 2,185,899 bp from pter )
and some psuedogenes are in:
plus strand (Start: 15,029,565 bp from pter End: up to 16,471,364 bp from pter)
Does it mean that my forward and reverse primers from the original gene can't amplify these psuedogenes?
thanks
mehr
thanks Devon