Team:Baltimore/Notebook
From 2011.igem.org
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Run through for abstract and scheduling: | Run through for abstract and scheduling: | ||
+ | |||
Start with plasmid with taq gene and pst1 gene | Start with plasmid with taq gene and pst1 gene | ||
+ | |||
Remove pst1 site from taq coding sequence | 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 | 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) | 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 | Make taq protein | ||
+ | |||
Compare it to other enzymes and make sure it works | Compare it to other enzymes and make sure it works |
Revision as of 02:12, 8 July 2011
This is a template page. READ THESE INSTRUCTIONS.
You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples HERE.
You MUST have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page. PLEASE keep all of your pages within your teams namespace.
You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. | File:Baltimore logo.png 200px |
Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs) | File:Baltimore team.png Your team picture |
Team Example |
Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
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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