FAQs
Get the most out of your enzyme with our documents, guides, brochures and tools
FAQs
Browse our frequently asked technical questions by selecting a product below.
How can high Ct’s in the qPCR step or low PCR product on the gel after a RT step (PCR/qPCR problem) be alleviated?
Perform gel electrophoresis to verify the amount of cDNA product. If the amount of cDNA product after the RT step is not sufficient, there might be a problem with RTase inhibiting Taq1. Try solving this by diluting the cDNA product 10-20x before adding 2.5 μL of this diluted reaction to 20 μL of total reaction volume. Adjust accordingly if the volumes are different. Alternatively, try diluting the enzyme itself before the RT step by carrying out 5x, 10x, 20x and 40x dilutions. These two procedures can also be combined in case further fine tuning is required to find the optimal result.
1 Suslov, O. & Steindler, D. A. PCR inhibition by reverse transcriptase leads to an overestimation of amplification efficiency. Nucleic Acids Res 33, e181, doi:10.1093/nar/gni176 (2005).
