Jan 21, 2016

Public workspacePSS2 PCR Protocol

DOI
dx.doi.org/10.17504/protocols.io.dfm3k5
  • Mel Duhaime1
  • 1Matthew Sullivan Lab, University of Arizona/Ohio State University
  • VERVE Net
  • Sullivan Lab
Verve Team
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DOI: dx.doi.org/10.17504/protocols.io.dfm3k5
Protocol Citation: Mel Duhaime 2016. PSS2 PCR Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.dfm3k5
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: July 16, 2015
Last Modified: November 09, 2017
Protocol Integer ID: 1229
Guidelines
Primers:

attP_F502: CTC CTC AAC TGC CAG GAC TC (PSS2 target 502bp upstream of att: 90315-90334)
attP_R141: CCC AACAAG CTC TCA GGA AG (PSS2 target 141bp downstream of att rev: 91010-91029)

Reaction Mix

ReagentAmt.Final Conc.
Water18.25μl
10x Buffer (no Mg)2.5μl=1x
25mM MgCl21.5μl=1.5mM
dNTP mix, 10mM each0.5μl=200μM ea.
Primer F502, 10μm0.5μl=0.2μM
Primer R141, 10mM0.5μl=0.2μM
Sigma Taq 5U/μl0.25μl=0.05U/μl
DNA (viral lysates)1.0μl
Total25μl


Cycling

95°C3 min
95°C15sec
35 cycles

60°C30sec
72°C30sec
72°C10min
15°CHold