FAQs
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FAQs
Browse our frequently asked technical questions by selecting a product below.
What is the error rate of the RTase in qPCRBIO cDNA Synthesis Kit?
Like the great majority of the RTases available on the market UltraScript Reverse Transcriptase, as well as all other RTases we offer, are derived from wild-type Moloney Murine Leukemia Virus (MMLV) RTase and have an error rate of 1×10-4 errors/bp1. This also applies to RTases derived from Avian Myeloblastosis Virus (AMV).
Running the reaction at a higher temperature increases fidelity because it destabilises mismatched base pairs2. If low fidelity is required adding manganese will make RTases highly mutagenic and could increase their speed3.
1 Yasukawa, K. et al. Next-generation sequencing-based analysis of reverse transcriptase fidelity. Biochem Biophys Res Commun 492, 147-153, doi:10.1016/j.bbrc.2017.07.169 (2017).
2 Malboeuf, C. M., Isaacs, S. J., Tran, N. H. & Kim, B. Thermal effects on reverse transcription: improvement of accuracy and processivity in cDNA synthesis. Biotechniques 30, 1074-1078, 1080, 1082, passim, doi:10.2144/01305rr06 (2001).
3 Cadwell, R. C. & Joyce, G. F. Randomization of genes by PCR mutagenesis. PCR Methods Appl 2, 28-33 (1992).
