Blue:RNA-Seq Experiments:05202013: Difference between revisions

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(Created page with "'''Library:''' UHRR 05162013 '''Purpose:''' Determine optimal conditions for Phusion HF and Advantage PCR reactions for initial library expansion of limiting DNA template (25...")
 
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Stage 1: 95C 15s --> 58C 30s --> 68C 60s {6 cycles}
Stage 1: 95C 15s --> 58C 30s --> 68C 60s {6 cycles}
Stage 2: 95C 15s --> 65C 30s --> 68C 60s {8-14 cycles}
Stage 2: 95C 15s --> 65C 30s --> 68C 60s {8-14 cycles}
[[File:05202013.pdf]]

Revision as of 00:14, 22 May 2013

Library: UHRR 05162013

Purpose: Determine optimal conditions for Phusion HF and Advantage 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}


File:05202013.pdf