Is DNase necessary for primers designed on the exon-exon boundary??
Posted 05 January 2011 - 05:51 PM
I wanted to know if genomic DNA contamination is a problem even if the primer itself designed on the exon-exon boundary? Since the gDNA wouldn't amplify at all if they were designed that way then should I forget about doing DNase treatment?
Posted 05 January 2011 - 05:58 PM
My two cents
Posted 11 January 2011 - 08:05 PM
If you do regular RT-PCR using such primers, DNA contamination can be found by the existence of a high sized band from the DNA. Despite that, If your purpose is to compare gene expression by judging the intensity of the bands and if the DNA contamination is serious, the amplification from the DNA may interfere with cDNA amplification and makes your interpretation of gene expression level inaccurate.