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Team:UIUC-Illinois/Notebook/Protocols/Bootcamp
From 2012.igem.org
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<center><h1>Bootcamp Protocols</h1></center> | <center><h1>Bootcamp Protocols</h1></center> | ||
| - | <h2>Making LB for plates</h2 | + | <h2>Making LB for plates</h2> <br/> |
| - | + | To make 1 Liter of LB: <br/><br/> | |
1. Always use a 10:5:5 ratio of Tryptone to Yeast Extract to NaCl. <br/><br/> | 1. Always use a 10:5:5 ratio of Tryptone to Yeast Extract to NaCl. <br/><br/> | ||
2. Add dry ingredients first. <br/><br/> | 2. Add dry ingredients first. <br/><br/> | ||
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6. Pour 25 mL on each plate (just enough to cover the bottom). <br/><br/> | 6. Pour 25 mL on each plate (just enough to cover the bottom). <br/><br/> | ||
| - | <h2>Making Liquid Media </h2 | + | <h2>Making Liquid Media </h2> <br/> |
Always use a 5-2.5-2.5 ratio of Tryptone to Yeast Extract to NaCl | Always use a 5-2.5-2.5 ratio of Tryptone to Yeast Extract to NaCl | ||
Add dry ingredients first, then add MilliQ water | Add dry ingredients first, then add MilliQ water | ||
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Autoclave by total volume | Autoclave by total volume | ||
| - | <h2>Making Glycerol Stock </h2 | + | <h2>Making Glycerol Stock </h2> <br/> |
To make 400 mL of 10% glycerol you will need:<br/><br/> | To make 400 mL of 10% glycerol you will need:<br/><br/> | ||
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4. Don’t autoclave! <br/><br/> | 4. Don’t autoclave! <br/><br/> | ||
| - | <h2>Making ddH2O</h2 | + | <h2>Making ddH2O</h2> <br/> |
- Use the small bottles. <br/><br/> | - Use the small bottles. <br/><br/> | ||
- Autoclave water by total volume. <br/><br/> | - Autoclave water by total volume. <br/><br/> | ||
| - | <h2>Autoclaving </h2 | + | <h2>Autoclaving </h2><br/> |
- Robert is the source of official help on all things about the Autoclave. <br/><br/> | - Robert is the source of official help on all things about the Autoclave. <br/><br/> | ||
1. Look at the Betastar for settings. <br/><br/> | 1. Look at the Betastar for settings. <br/><br/> | ||
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Use the autoclave tape and make sure it has changed colour by the end of the cycle. <br/><br/> | Use the autoclave tape and make sure it has changed colour by the end of the cycle. <br/><br/> | ||
| - | <h2>Ligation </h2 | + | <h2>Ligation </h2><br/> |
- We used the <a href="http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf">Ginkgo bioworks protocol</a> <br/><br/> | - We used the <a href="http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf">Ginkgo bioworks protocol</a> <br/><br/> | ||
- Control: 2 uL of Circular PSB1C3 plasmid, 13 uL ddH2O, 2 uL T4 ligase buffer, 1 uL T4 ligase <br/><br/> | - Control: 2 uL of Circular PSB1C3 plasmid, 13 uL ddH2O, 2 uL T4 ligase buffer, 1 uL T4 ligase <br/><br/> | ||
Revision as of 19:12, 1 June 2012
