What are the critical factors which hinder/inhibit the cDNA synthesis ?

14 December 2016 1 1K Report

Dear friends ,

I isolated the total RNA from Coleus stem and the RNA looked good on gel (below). I made the cDNA from the RNA but when i checked PCR with Actin(250bp) and gene specific primer, it showed no amplification.

plz tell me what are the possible steps where i may be faulty

Sheikh Gulfam

thanks,

Gene specific primer showed a slight band but actin was completely absent

plz see gel picture below

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