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Team:Washington/HOMEPAGE PROTOCOL
From 2013.igem.org
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| - | <u>Cloning Protocols Workflow</u> | + | <b><u>Cloning Protocols Workflow</u></b> |
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| - | Tips and Etiquette: | + | <b>Tips and Etiquette:</b> |
<ul> | <ul> | ||
<li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">General Basic Protocol Information</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BASIC_PROTOCOL">General Basic Protocol Information</a></li> | ||
| - | <li><a href = "https://2013.igem.org/Team:Washington/ | + | <li><a href = "https://2013.igem.org/Team:Washington/HOW_TO_LABEL_EVERYTHING">How to Label EVERYTHING</a></li> |
</ul> | </ul> | ||
Keep notebooks up-to-date | Keep notebooks up-to-date | ||
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<br> | <br> | ||
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| + | <p> | ||
| + | <b>Workflow:</b> | ||
| + | <ul> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/PRIMER_DESIGN">Primer Design</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/STREAKING_A_PLATE">Streaking a Plate for Isolation</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/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> | ||
| + | </p> | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <p> | ||
| + | <b>Media Preparation:</b> | ||
| + | <ul> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/COMPETENT_CELL_STOCKS">Competent Cell Stocks</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/LB_AND_TB_MEDIA">LB and TB Media</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/POURING_AGAR_PLATES">Pouring Agar Plates</a></li> | ||
| + | </ul> | ||
| + | </p> | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <p> | ||
| + | <b>Instruments:</b> | ||
| + | <ul> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/NANODROP_PROTOCOL">Nanodrop Protocol</a></li> | ||
| + | </ul> | ||
| + | </p> | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <p> | ||
| + | <b>See also:</b> | ||
| + | <ul> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/GENERAL_BIOTECHNOLOGY_INFORMATION">General Biotechnology Information</a></li> | ||
| + | <li><a href = "https://2013.igem.org/Team:Washington/SOS_LAB_PROTOCOLS">SOS Lab Protocols</a></li> | ||
| + | <li><a href = "http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf">http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf</a></li> | ||
| + | </ul> | ||
| + | </p> | ||
</body> | </body> | ||
</html> | </html> | ||
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
