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Team:Washington/Protocols
From 2013.igem.org
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| - | <b><u>Cloning Protocols | + | <b><u>Cloning Protocols</u></b> |
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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/General_Cloning_Strategy">General Cloning Strategy</a></li> | <li><a href = "https://2013.igem.org/Team:Washington/General_Cloning_Strategy">General Cloning Strategy</a></li> | ||
| + | </ul> | ||
| + | <b>Workflow:</b> | ||
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| + | <ul> | ||
<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 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_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> | ||
Revision as of 21:32, 27 September 2013
Cloning Protocols Workflow:
- 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)
- 9) Making Glycerol Frozen Stocks
Light Sensing Protocols
Light Sensing Media:
5X M9 Salts M9 Media + Casamino Acid Recipe M9 + Casamino Acid Plate Light Testing:General Protocol 96-Well Plate Assay 60 mm Petri Dish Assay Fluorescent Analysis Protocol GFP Expression on Agar Plates Light Intensity Test Blinking Test Setting Up The App:E.colight Setup App Testing:Bleed-Through Test
