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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 | ||
| - | + | ===July, Week 2=== | |
| - | 7-12-11 | + | ====7-12-11==== |
Made one agarose gel 1% | Made one agarose gel 1% | ||
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--[[User:Mduley|Mduley]] 19:19, 12 July 2011 (CDT) | --[[User:Mduley|Mduley]] 19:19, 12 July 2011 (CDT) | ||
| - | 7-13-2011 | + | ====7-13-2011==== |
Retrieved gel from cold room and covered with 1x TAE in gel box and heated it (by running the gel with nothing in it) | Retrieved gel from cold room and covered with 1x TAE in gel box and heated it (by running the gel with nothing in it) | ||
Revision as of 17:24, 14 July 2011
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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
July, Week 2
7-12-11
Made one agarose gel 1% Used 0.5g Agarose and 50 mLs x TAE Poured the gel in the cold room and covered with a papre towel labled iGEM --Mduley 19:19, 12 July 2011 (CDT)
7-13-2011
Retrieved gel from cold room and covered with 1x TAE in gel box and heated it (by running the gel with nothing in it)
Prepared Mutagenesis PCR products from last Thursday to be run on Gel
The tubes I made up came from the following:
A = Green/ Blue tubes conatained Dr. Burkett's dNTP's B = Orange tubes contain Dr. Goode's dNTP's
I prepared 8 tubes as follows:
CA = Pink tube contains 10 microliters of the Control A sample and 2 microliters of loading dye
CB = Blue tube contains 10 microliters of the Control B sample and 2 microliters of loading dye
Rxn 1 A & B = Pink A tube contains 10 microliters of the RXN 1 A sample and 2 microliters of loading dye
Blue B tube contains 10 microliters of the RXN 1 B sample and 2 microliters of loading dye
Rxn 4 A & B = Pink A tube contains 10 microliters of the RXN 4 A sample and 2 microliters of loading dye
Blue B tube contains 10 microliters of the RXN 4 B sample and 2 microliters of loading dye
Rxn 5 A & B = Pink A tube contains 10 microliters of the RXN 5 A sample and 2 microliters of loading dye
Blue B tube contains 10 microliters of the RXN 5 B sample and 2 microliters of loading dye
These 8 tubs are located in the freezer and are ready to be run on the gel. They are in a yellow rack with a pink taped labeled with todays date and iGEM, mutagenesis PCR for gel.
Dr. Goode requested that the gel lanes be filled as follows
1. Ladder 2. CA 3. CB 4. R1A 5. R1B 6. Ladder 7. R4A 8. R4B 9. R5A 10. R5B
--Mduley 18:00, 13 July 2011 (CDT)
