Real-time PCR as is more sensitive than RT PCR but is it still possible to get positive reaction from RT PCR while negative in real-time PCR.
Actually RT PCR was done earlier in which I got positive rxn. but when same samples were put to Real time PCR after a gap of few months negative results were found in few samples. Could it be due to the degradation of mRna in these samples as less amount of that might be present??
again do RT PCR, to check the integrity of your samples and primers.
i don,t understand r u using RNA or cDNA, if cDNA, cDNA not degrade on -20C. conform your Kit for Real time that not exposed in light, temperature or reaction condition were same as RT-PCR.
thereafter, i again used mRNA for cdna synthesis and found few samples negative.
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