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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/OVERNIGHT_CULTURES">Overnight Cultures</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/OVERNIGHT_CULTURES">Overnight Cultures</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 = "">PCR GoTag</a> - product (30-200ng/ ul) Check on gel PCR Purification and/or <a href = "">Dpnl Digest</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/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> | ||
Latest revision as of 02:11, 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 Miniprep (stocks can be made from this culture - add 1ml extra)
- 8) DNA Sequencing
- 9) Making Glycerol Frozen Stocks
Media Preparation:
Instruments:
See also:
- General Biotechnology Information
- SOS Lab Protocols
- http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf
