Team:Kyoto/Medium

From 2011.igem.org

Contents

Medium

Standard medium for drosophila

  1. Stir dry yeast 20g and agarose 3.5~5.0g with about two-thirds of water 500mL. Then, autoclave it.
  2. Stir corn flour 45g and glucose 50g with the remaining water.
  3. Stir 1 and 2, then autoclave it again.
  4. after autoclave, add propionic acid 1.5mL and 10 % p-hydroxybenzoate in 70 % Eternol 5mL into it.

Standard medium for drosophila

M9 medium

  1. Stir Na2HPO4 6 g, KH2PO4 3 g, NaCl 0.5 g and NH4Cl 1 g with water 1 l.
  2. After autoclave, add 10 ml filter sterilized 100 mM MgSO4, 20 % glucose, 10 mM CaCl2, 100 mM thiamine-HCl.
  3. If you need, add 10 ml filter sterilized 20 % casamino acid.

LB medium

  1. Stir Tryptone 20 g, Yeast extract 10 g and NaCl 10 g with water 200ml.
  2. If you make LB plates, add agar 10 g.
  3. Autoclave.

SOB medium

  1. Stir Tryptone 20g and Yeast extract 5g with water.
  2. Add 2 ml 5 M NaCl and 840 ul 3 M KCl and add water up to 1 l.
  3. After autoclave, add 10 ml filter sterilized 1 M MgSO4 and 1 M MgCl2.

SOC medium

  1. Add 2 M glucose 1 ml to 100 ml SOB.

Buffer TB

  1. Stir PIPES and CaCl2・2H2O with 100 ml water.
  2. Add 8.315 ml 3 M KCl.
  3. Add KOH and adjust pH 6.8.
  4. Add MnCl2・4H2O.
  5. Add water up to 200 ml.
  6. Filter sterilize

DNS reaegnt

  1. Add 1% DNS 88ml and Rochelle salt 25.5 g to 4.5% NaOH 30 ml (A solution)
  2. Add Phenol 1 g and water 7.8 ml to 10% NaOH 2.2 ml (B solution)
  3. Add NaHCO3 6.9g to B solution 6.9 ml
  4. Add A solution 118ml to B solution 6.9 ml
  5. Leave 2 days
  6. Store in brown bottle
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