IGEM:British Columbia/Protocols/Amplifying BioBrick Parts
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Amplifying Biobrick Parts
Materials
- 2009 Spring Distribution plates
- Competent cells
- sdH2O
- LB agar plates (with appropriate antibiotics)
- LB broth (with appropriate antibiotics)
Equipment
- miniprep kits (1 prep/part)
Method
- With a pipette tip, punch a hole through the foil cover into the well corresponding to the part you want. Make sure that the plate is properly oriented
- Resuspend in a PCR tube with 15uL of sdH2O (by pipetting in and out of the water).
- Transform competent cells with 1uL of the resuspended DNA, plate bacteria with the appropriate antibiotic, and grow overnight.
- Pick a single colony and inoculate ~3.5-6mL of medium (with the correct antibiotic) and grow overnight (16-18 hours).
- Make a glycerol stock using 0.75mL of culture.
- Miniprep the DNA using ~2.5-5mL of culture
- Label well and store at -4°C.
