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Team:Washington/HOMEPAGE PROTOCOL
From 2013.igem.org
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<li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_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_BASIC_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_BASIC_PROTOCOL">4) Agarose Gel Electrophoresis</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">4) Agarose Gel Electrophoresis</a></li> | ||
| - | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">5) General Ligation Protocol</a></li> | + | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_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/GENERAL_BASIC_PROTOCOL">6) Heat shock/chemical competent transformation</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">6) Heat shock/chemical competent transformation</a></li> | ||
<li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">7) Colony PCR with Green taq</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">7) Colony PCR with Green taq</a></li> | ||
Revision as of 01:52, 7 September 2013
Tips and Etiquette:
Keep notebooks up-to-dateWorkflow:
- Primer Design
- Streaking a Plate for Isolation
- Overnight Cultures
- 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
- 8) DNA Sequencing
- 9) Making Glycerol Frozen Stocks
