When performing a qPCR one usually must dilute the first strand reaction before going into the PCR part. I'm trying to do both reactions at the same time in the same tube so I have to load all the reagents for both reactions at once. I'm having some success. I'm wondering, however, if anyone out there knows what component of the cDNA synthesis (buffer, MMLV, DTT, primers, nucleotides, excessive cDNA product) has an inhibitory effect upon the PCR portion of a qPCR?
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Which cDNA synthesis reagents/products affect qPCR?
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