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From 2011.igem.org

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Revision as of 21:44, 4 October 2011

Contents 1 Medium for drosophila 2 M9 medium 3 LB medium 4 SOB medium 5 SOC medium 6 Buffer TB 7 DNS reaegnt

Medium for drosophila

1. Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it.
2. Stir corn flour and glucose with the remaining water.
3. Stir 1 and 2, then autoclave it again.
4. after autoclave, add propionic acid and 10 % p-hydroxybenzoate in 70 % Eternol into it.

[http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html 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