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Team:Baltimore/Notebook
From 2011.igem.org
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Compare it to other enzymes and make sure it works | Compare it to other enzymes and make sure it works | ||
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| + | 7-12-11 | ||
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| + | Made one agarose gel 1% | ||
| + | --[[User:Mduley|Mduley]] 19:19, 12 July 2011 (CDT) | ||
Revision as of 00:19, 13 July 2011
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Notebook
Run through for abstract and scheduling:
Start with plasmid with taq gene and pst1 gene
Remove pst1 site from taq coding sequence
- In order to remove the restriction site, do site directed mutagenesis (1day)
- PCR
- Digestion
- Transformation
- Screening (1day)
- Dilute DNA
- Colony PCR individually
- Digestion of PCR product with pst1
- Run gel ~2500bp
Add biobrick prefix/suffix to taq coding sequence
- PCR and add the prefix and suffix as primers (one prefix has an extra AG-make sure to use the correct one) (1-2 days)
- Run gel
- Cut out of gel
- Cut with restriction enzyme
- Clean up DNA
- Ligation with vector (could be vector with the terminator sequence, promoter and RBS)
- Transformation
Add a promoter, transcriptional terminator, ribosome binding site (RBS)
- Screen colonies (1 day)
- Colony PCR
- Restriction Digestion
- Clean up DNA
- Sequence (1 day)
Make taq protein
Compare it to other enzymes and make sure it works
7-12-11
Made one agarose gel 1% --Mduley 19:19, 12 July 2011 (CDT)
