Primers amplify genomic DNA but not cDNA
Posted 01 August 2012 - 10:27 AM
Any thoughts anyone, pretty soon I'm going to be banging my head against the desk in pure frustration as I'm running out of options to try.
Posted 01 August 2012 - 01:30 PM
I'm affraid from "total RNA" kits you won't get unspliced RNA in sufficient amounts, if that was true, people would get get two types of bands in their intron spanning PCR assays even when having no DNA contamination.
Your first primer, it's probably in intronic region, right? Are the other primers in introns too?
Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.
I never trust anything that can't be doubted.
'Normal' is a dryer setting. - Elizabeth Moon
Posted 02 August 2012 - 04:12 AM