IGEM:British Columbia/Protocols/Amplifying BioBrick Parts

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Revision as of 14:54, 7 August 2009 by Heather Kempthorne (talk | contribs) (New page: ==Amplifying Biobrick Parts== ===Materials=== * 2009 Spring Distribution plates * Competent cells * sdH2O * LB agar plates (with appropriate antibiotics) * LB broth (with appropriate anti...)
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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.

4. Pick a single colony and inoculate ~3.5-6mL of medium (with the correct antibiotic) and grow overnight (16-18 hours). 5. Make a glycerol stock using 0.75mL of culture. 6. Miniprep the DNA using ~2.5-5mL of culture 7. Label well and store at -4°C.