From 2014.igem.org
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| - | <a name="btrans"><h2>Bacterial Transformation</h2> | + | <a name="btrans"></a><h2>Bacterial Transformation</h2> |
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| - | <a name="cpcr"><h2>Colony PCR</h2> | + | <a name="cpcr"></a><h2>Colony PCR</h2> |
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| - | <a name="gel"><h2>Gel Extraction</h2> | + | <a name="gel"></a><h2>Gel Extraction</h2> |
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| - | <a name="mini"><h2>Miniprep</h2> | + | <a name="mini"></a><h2>Miniprep</h2> |
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| - | <a name="pcrpure"><h2>PCR Purification</h2> | + | <a name="pcrpure"></a><h2>PCR Purification</h2> |
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| - | <a name="pcr"><h2>PCR Reaction</h2> | + | <a name="pcr"></a><h2>PCR Reaction</h2> |
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| - | <a name="tdtheat"><h2>TdT Heat Inactivation</h2> | + | <a name="tdtheat"></a><h2>TdT Heat Inactivation</h2> |
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| - | <a name="tdtbase"><h2>TdT Nucleotide Addition</h2> | + | <a name="tdtbase"></a><h2>TdT Nucleotide Addition</h2> |
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| - | <a name="gene"><h2>Yeast Gene Knockout</h2> | + | <a name="gene"></a><h2>Yeast Gene Knockout</h2> |
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| - | <a name="mate"><h2>Yeast Mating</h2> | + | <a name="mate"></a><h2>Yeast Mating</h2> |
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| - | <a name="spore"><h2>Yeast Sporulation/Zymolyase Treatment</h2> | + | <a name="spore"></a><h2>Yeast Sporulation/Zymolyase Treatment</h2> |
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Revision as of 23:27, 14 October 2014
Cooper Union 2014 iGEM
Common Laboratory Protocols
Bacterial Transformation
Colony PCR
Gel Extraction
Miniprep
PCR Purification
PCR Reaction
Restriction Enzyme Digest
- Materials
- 10x Buffer (Compatible to the enzyme)
- Enzyme
- BSA - diluted to 10x
- ddH2O
- DNA - up to 1μg per reaction.
Procedure
Verify all times and temperatures for specific enzyme prior to use. This is general protocol.
- Prewarm 2 water baths to 37°C and 70°C
- Thaw buffers, diluted BSA. Keep all reagents on ice
- Set up reactions based on table below.
| Reagent | Volume | Example |
| Reaction Buffer | 1/10 total volume | 2μL |
| Diluted BSA (10x) | 1/10 total volume | 2μL |
| DNA | calculate for target ng | 3.7μL |
| ddH2O | volume up to (total volume - 1μL) | 11.3μL |
| Enzyme | generally 1μL | 1μL |
| Total Volume | | 20uL |
- Incubate 60min 37°C.
- Heat inactive 70°C for 15 min.
- Store in -20°C or use immediately.
Top of Page
TdT Heat Inactivation
TdT Nucleotide Addition
Yeast Gene Knockout
Yeast Mating
Yeast Sporulation/Zymolyase Treatment
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