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Team:Baltimore/Notebook
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Contents |
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
Calendar
July
Week 2: Tuesday, July 12; Wednesday, July 13; Thursday, July 14; Friday, July 15; Saturday, July 16; Sunday, July 17;
Week 3: Tuesday, July 19; Wednesday, July 20; Thursday, July 21; Friday, July 22; Saturday, July 23; Sunday, July 24; took 1 ul of each of products after npt1 digestion done last thursday, put on ice, added 25 ul of l to each, shocked at 42C for 30 seconnds, placed back on ice for 1 half hour, put onto amp plates into warm room. On Monday, got plates out of warm room, put into cold room late (4:00 pm)
Week 4: Tuesday, July 26; Wednesday, July 27; Thursday, July 28; Friday, July 29; Saturday, July 30; Sunday, July 31;
August
Week 1: Tuesday, August 2; Wednesday, August 3; Thursday, August 4; Friday, August 5; Saturday, August 6; Sunday, August 7;
Week 2: Tuesday, August 9; Wednesday, August 10; Thursday, August 11; Friday, August 12; Saturday, August 13; Sunday, August 14;
Week 3: Tuesday, August 16; Wednesday, August 17; Thursday, August 18; Friday, August 19; Saturday, August 20; Sunday, August 21;
Week 4: Tuesday, August 23; Wednesday, August 24; Thursday, August 25; Friday, August 26; Saturday, August 27; Sunday, August 28;
