Blue:RNA-Seq Experiments:05202013: Difference between revisions
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'''Library:''' UHRR 05162013 | '''Library:''' UHRR 05162013 | ||
'''Purpose:''' | '''Purpose:''' Determined optimal conditions for Phusion HF and Advantage (Clontech) PCR reactions for initial library expansion of limiting DNA template (250pg Library) | ||
'''Outcome:''' Phusion HF worked slightly better than Advantage and had less ssDNA amplification. Both Phusion and Advantage worked better than Sybr | '''Outcome:''' Phusion HF worked slightly better than Advantage and had less ssDNA amplification. Both Phusion and Advantage worked better than Sybr |
Latest revision as of 00:18, 22 May 2013
Library: UHRR 05162013
Purpose: Determined optimal conditions for Phusion HF and Advantage (Clontech) PCR reactions for initial library expansion of limiting DNA template (250pg Library)
Outcome: Phusion HF worked slightly better than Advantage and had less ssDNA amplification. Both Phusion and Advantage worked better than Sybr
Phusion:
Stage 1: 95-98C 10s --> 58C 20s --> 72C 30s {6 cycles} Stage 2: 95-98C 10s --> 72C 20s {8-14 cycles}
Advantage:
Stage 1: 95C 15s --> 58C 30s --> 68C 60s {6 cycles} Stage 2: 95C 15s --> 65C 30s --> 68C 60s {8-14 cycles}