IGEM:British Columbia/Protocols/Amplifying BioBrick Parts

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Contents

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.
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