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_23
Snippet: As an alternative to the MUTsl-A7 construct whose potential for 'model' stem-loop structure formation is abolished by base substitution, we employed an RNA antisense strategy to decrease the potential for model stemloop structure formation in the wtSL-A7 template. The result showed that presence of a 10 nt antisense RNA (anti-5 stem), complementary to the RNA sequence 9 nt 5 to the 7A's, modestly decreases one base addition and enhances base omis.....
Document: As an alternative to the MUTsl-A7 construct whose potential for 'model' stem-loop structure formation is abolished by base substitution, we employed an RNA antisense strategy to decrease the potential for model stemloop structure formation in the wtSL-A7 template. The result showed that presence of a 10 nt antisense RNA (anti-5 stem), complementary to the RNA sequence 9 nt 5 to the 7A's, modestly decreases one base addition and enhances base omission. These experiments also showed that the efficiency and/or the directionality of the realignment are affected depending on the dNTP ratios (Supplementary Data and Figure S1 ).
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