I am using a Bio-Rad C1000 touch thermo cycler and I have a query regarding my PCR amplification protocol. I am doing long PCR amplifications with product size ~11 to 14kb. Now my products amplify perfectly when I run a temperature gradient between 550C – 590C and I get amplified products of the correct size for each temperature condition. However the PCR amplification fails to work if I run the block without a temperature gradient (i.e. at a set temperature of 550C)
I was wondering if this is a known problem for C1000 users and if there are any suggestions to get around this?