Team:Hawaii/Protocols/Solid growth media

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Contents

LB agar

Materials

  • 25 g LB broth mixture (Difco)
  • 15 g agar
  • 1 L ddH2O

Procedure

  1. Mix dry ingredients together with 1 L ddH2O using a magnetic stir bar.
  2. Autoclave to sterilize. Let cool to 55C.
  3. Pour 28 ml LB solution for thin plates; 50 ml for thick plates.

LB + amp100

Materials

Procedure

  1. Make LB agar as directed.
  2. When mixture is cooled to 55C, add 1 ml 1000X ampicilllin stock solution.
  3. Plate as usual.

LB+kan20+amp50

Materials

Procedure

  1. Make LB as directed.
  2. After cooling to 55C, add ampicillin and kanamycin.
  3. Plate as usual.

LB+sp100

Materials

Procedure

  1. Make LB as directed.
  2. Cool to 55C, then add spectinomycin.
  3. Plate as usual.

BG-11 agar

Materials

  • 940 ml nanopure H2O
  • 20 ml 50X thiosulfate (final conc. of 1mM)
  • 20 ml 50X TES buffer, pH 8.0 (final conc. of 10mM)
  • 20 ml 50X BG-11
  • 15 g agar

Procedure

  1. Combine agar and nanopure H2O. Stir mixture with a magnetic stir bar.
  2. Autoclave to sterilize. Cool to 55C.
  3. Add TES, thiosulfate, and BG-11
  4. Test pH (should be ~8.0).
  5. Plate 28 ml for thin plates; 50 mL for thick plates.

BG-11 + 5% LB

  • 1.25 g LB broth mixture (Difco)
  • 15 g agar
  • 940 ml nanopure H2O
  • 20 ml 50X BG-11
  • 20 ml 50X thiosulfate
  • 20 ml 50X TES buffer
  1. Combine LB and agar with the nanopure H2O.
  2. Autoclave to sterilize.
  3. Add TES, thiosulfate, and BG-11.
  4. Test pH (~8.0).
  5. Plate as usual.

BG-11+sp2.5+sm2.5

Materials

Procedure

  1. Make BG-11 agar as directed.
  2. After adding TES, thiosulfate, and BG-11, add spectinomycin and streptomycin.
  3. Plate as usual.