Team:Hawaii/Protocols/Solid growth media

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.

Materials

 * 1 L LB agar
 * 1 ml 1000X ampicillin stock solution

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.

Materials

 * 1 L LB agar
 * 0.5 mL 1000X ampicillin stock solution
 * 0.4 mL 1000X kanamycin stock solution

Procedure

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

Materials

 * 1 L LB agar
 * 1.0 mL 1000X spectinomycin stock solution

Procedure

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

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.

Materials

 * 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

Procedure

 * 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.

Materials

 * BG-11 agar
 * 0.15 ml 1000X spectinomycin stock solution
 * 0.3 ml 1000X streptomycin stock solution

Procedure

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