I am looking for some help on PCR. We are attempting to PCR
up a 3.5kb gene from human DNA, so far unsuccessfully.
Q1. Is it too ambitious to go for the entire 3.5kb with
one set of amplimers? What is the record size acheived
from total genomic DNA?
Q2. Our gene contains an intron with an Alu sequence in it.
Could this be the reason why we cannot get ful length product?
We have postulated that the alu sequences re-anneal very quickly
and prevent polymerase extension through this region...anybody
have any thoughts/observations on this?
Q3. If anyone out there has PCR'd for 3-4kb through an alu sequence,
can you tell me how you did it?!!!
Thanks.............Mike Dalrymple, CAGR, Edinburgh.