From 2011.igem.org

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]

Gene Assembly With Oligos

Computational Protein Design

Sequencing

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

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