Stimulation of reverse transcriptase generated cDNAs with specific indels by template RNA structure: retrotransposon, dNTP balance, RT-reagent usage

Penno, Christophe; Kumari, Romika; Baranov, Pavel V.; van Sinderen, Douwe; Atkins, John F.

The reverse transcriptase from WT Moloney Murine Leukemia virus (MuLV), the parent of SuperScriptâ„¢ III, plus the RTs from Avian Myeloblastosis virus (AMV) and from HIV-1 and HIV-2 were similarly tested with WTsl-A7, and mutSL-A7 under the nine dNTPs concentration conditions. In addition these RTs were tested with the WT, or mutated, 'model' stem-loop 5 adjacent to an A6 motif (wtSL-A6 and MUTsl-A6) under equimolar dNTP concentration condition. The results show a similar LPE product pattern indicating that at identical RNA template and dNTP concentration conditions, the different RT polymerases tested share a clear similar response to slippage directionality. However, for specific reaction conditions, they can show marked differences in their slippage propensity (Supplementary Results and Supplementary Figure S5) .