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