Author: Penno, Christophe; Kumari, Romika; Baranov, Pavel V.; van Sinderen, Douwe; Atkins, John F.
Title: Stimulation of reverse transcriptase generated cDNAs with specific indels by template RNA structure: retrotransposon, dNTP balance, RT-reagent usage Document date: 2017_9_29
ID: k4gtl2o7_8
Snippet: For the experiments included in that publication, the controls to distinguish whether indels in its product DNA derived from the initial DNA-dependent RNA polymerase step, or subsequently from product cDNA reverse transcriptase slippage, revealed no reverse transcriptase slippage. Follow-up work tested potential nascent RNA stemloop structure stimulation of slippage at runs of A shorter than 9, the minimal needed for efficient slippage at such mo.....
Document: For the experiments included in that publication, the controls to distinguish whether indels in its product DNA derived from the initial DNA-dependent RNA polymerase step, or subsequently from product cDNA reverse transcriptase slippage, revealed no reverse transcriptase slippage. Follow-up work tested potential nascent RNA stemloop structure stimulation of slippage at runs of A shorter than 9, the minimal needed for efficient slippage at such motifs. In this unpublished work, a significant proportion of the reverse transcriptase product of one 75 nt chemically synthesized RNA template with an inverted repeat with potential to form the 'model' stem-loop 5 adjacent to 8 A's, had an extra T. This control experiment prompted the present investigation of RNA template stemloop structure-mediated reverse transcriptase realignment.
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