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real time PCR-Primer question - (Nov/21/2006 )

what does it mean when you amplify the same gene with two different primers and you see two completely different cT values - shouldnt the values be similar, assuming the amplicon size is the same for both primers?



It might not similar due to different primer set with different amplification efficiency although amplicon size is the same.

On another hand, using polyT primer for reverse transcription might result in less signal for primer set target to 5' part of examined RNA, and using random primer for reverse transcription, might product less sequence contain whole 3' part of examined RNA.