Team:Washington/Protocols

From 2011.igem.org

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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