"
Team:Washington/Protocols
From 2013.igem.org
(Difference between revisions)
| Line 653: | Line 653: | ||
<ul> | <ul> | ||
<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/GENERAL_PCR_PROTOCOL">2) General PCR Protocol </a>(also see <a href = "https://2013.igem.org/Team:Washington/PCR_GOTAG">PCR GoTag</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/AGAROSE_GEL">4) Agarose Gel Electrophoresis</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_LIGATION_PROTOCOL">5) General Ligation Protocol</a> (Don’t forget background control plates) Heat Inactivation (optional - up to 10 fold increase) - 65° for 10 minutes</li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/HEAT_SHOCK_CHEM_TRANS">6) Heat shock/chemical competent transformation</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/COLONY_PCR_GREEN">7) Colony PCR with Green taq</a> Miniprep (stocks can be made from this culture - add 1ml extra)</li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/DNA_SEQUENCING">8) DNA Sequencing</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/MAKING_GLYCEROL_FROZEN_STOCKS">9) Making Glycerol Frozen Stocks</a></li> | ||
</ul> | </ul> | ||
</p> | </p> | ||
Revision as of 01:00, 25 September 2013
Workflow:
- iGEM Vector Information
- 1) Isolation of Plasmid DNA (miniprep)
- 2) General PCR Protocol (also see PCR GoTag - 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
