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Team:Washington/Protocols
From 2013.igem.org
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<li><a href = "https://2013.igem.org/Team:Washington/iGEMVectors">iGEM Vector Information</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/iGEMVectors">iGEM Vector Information</a></li> | ||
<li><a href = "https://2013.igem.org/Team:Washington/ISOLATION_OF_PLASMID">1) Isolation of Plasmid DNA (miniprep)</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/ISOLATION_OF_PLASMID">1) Isolation of Plasmid DNA (miniprep)</a></li> | ||
| - | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_PCR_PROTOCOL">2) General PCR Protocol </a>(also see <a href = "https://2013.igem.org/Team:Washington/PCR_GOTAG">PCR | + | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_PCR_PROTOCOL">2) General PCR Protocol </a>(also see <a href = "https://2013.igem.org/Team:Washington/PCR_GOTAG">PCR GoTaq</a> - product (30-200ng/ ul) Check on gel PCR Purification and/or <a href = "https://2013.igem.org/Team:Washington/DPNL_DIGEST">Dpnl Digest</a></li> |
<li><a href = "https://2013.igem.org/Team:Washington/GENERAL_DIGESTION_PROTOCOL">3) General Digestion Protocol</a> Check on gel PCR Purification or Heat Inactivation (check enzyme temp and time, usually 80C for 20min)</li> | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_DIGESTION_PROTOCOL">3) General Digestion Protocol</a> Check on gel PCR Purification or Heat Inactivation (check enzyme temp and time, usually 80C for 20min)</li> | ||
<li><a href = "https://2013.igem.org/Team:Washington/AGAROSE_GEL">4) Agarose Gel Electrophoresis</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/AGAROSE_GEL">4) Agarose Gel Electrophoresis</a></li> | ||
Revision as of 01:25, 25 September 2013
Workflow:
- iGEM Vector Information
- 1) Isolation of Plasmid DNA (miniprep)
- 2) General PCR Protocol (also see PCR GoTaq - product (30-200ng/ ul) Check on gel PCR Purification and/or Dpnl Digest
- 3) General Digestion Protocol Check on gel PCR Purification or Heat Inactivation (check enzyme temp and time, usually 80C for 20min)
- 4) Agarose Gel Electrophoresis
- 5) General Ligation Protocol (Don’t forget background control plates) Heat Inactivation (optional - up to 10 fold increase) - 65° for 10 minutes
- 6) Heat shock/chemical competent transformation
- 7) Colony PCR with Green taq Miniprep (stocks can be made from this culture - add 1ml extra)
- 8) DNA Sequencing
- 9) Making Glycerol Frozen Stocks
