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_16
Snippet: Each specific cDNA was amplified using the corresponding set of forward and reverse primers (Supplementary Table S1 ). Standard PCR reactions were 50 l volume and contained: 1× Thermo buffer (Biolabs), 2 l cDNA or 4 nM DNA oligo, 200 M each dNTP (Biolabs), 500 nM each specific primer, and 0.8 unit Taq DNA polymerase (Biolabs). The PCR cycle was: denaturation at 94 • C for 5 min, then 25 cycles of denaturation at 94 • C for 30 s, annealing a.....
Document: Each specific cDNA was amplified using the corresponding set of forward and reverse primers (Supplementary Table S1 ). Standard PCR reactions were 50 l volume and contained: 1× Thermo buffer (Biolabs), 2 l cDNA or 4 nM DNA oligo, 200 M each dNTP (Biolabs), 500 nM each specific primer, and 0.8 unit Taq DNA polymerase (Biolabs). The PCR cycle was: denaturation at 94 • C for 5 min, then 25 cycles of denaturation at 94 • C for 30 s, annealing at 52 • C for 30 s and elongation at 72 • C for 30 s. This was followed by a final elongation at 72 • C for 1 min.
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