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Team:Washington/Protocols
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[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works] | [http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works] | ||
| - | + | [https://2011.igem.org/Team:Washington/Protocols/expression_purification Standard 1L Expression Purification] | |
[https://2011.igem.org/Team:Washington/Protocols/gene_assembly Gene Assembly With Oligos] | [https://2011.igem.org/Team:Washington/Protocols/gene_assembly Gene Assembly With Oligos] | ||
Revision as of 22:25, 13 September 2011
General Protocols
General Transformation Protocol
[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html Overview of how Kunkel Mutagensis works]
Standard 1L Expression Purification
Make It: Diesel Production Protocols
Alkane Biosynthesis media and extraction
Break It: Gluten Destruction Protocols
Small Scale (50mL) Protein Expression and Purification
Make It: Magnetosome Protocols
Gibson Vectors (pGB) protocols
Check out or add wiki design tools here: Wiki Design
Colony PCR with Green tag
Master mix(7ul):
1ul 10uM forword primer
1ul 10uM reverse Primer
5ul 2x Green tag
Cell water(3ul): Pick one colony from the plate and mix with 10ul of ddH2O to make 10ul of cell water
Reaction = Master mix(7ul) + Cell water(3ul) = 10ul per tube
Use program "Colony" & change the extention time (1min per kb)
Heat Shock Transformation
2 ul ligation
20 ul cells
Ice 20 minutes
Heat shock at 42C for 1 minute
Ice 2 minutes
Prepare 200 ul of TB (no anti) and transformed cells in culture tube
Incubate at 37C for 1 hour
Plate cells
