Team:Washington/Protocols

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

[http://www.bio.davidson.edu/Courses/molbio/kunkel/kunkel.html 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

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