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Protocol

Medium for drosophila

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

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

M9 medium

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

LB medium

Materials	Methods
water : 1 L Tryptone: 10 g Yeast extract : 5 g NaCl : 10 g Agar : 15g	Stir Tryptone, Yeast extract and NaCl with water. If you make LB plates, add agar. Autoclave.

SOB medium

Materials	Methods
water : 100 mL Tryptone: 2 g Yeast extract : 0.5 g 5 M NaCl : 200 ul 3 M KCl : 84 ul 1 M MgSO4 : 1 ml 1 M MgCl2 : 1 ml	Stir Tryptone and Yeast extract with water. Add 200 ul 5 M NaCl and 84 ul 3 M KCl. After autoclave, add 1 ml filter sterilized 10 mM MgSO4 and 10 mM MgCl2.

SOC medium

Materials	Methods
SOB : 100 ml 2 M glucose: 1 ml	Add 2 M glucose to 100 ml SOB.

Buffer TB

Materials	Methods
water : 200 ml PIPES : 3 g CaCl2・2H2O : 0.22 g 3 M KCl : 8.315 ml KOH MnCl2・4H2O : 1.09 g	Stir PIPES and CaCl2・2H2O with 100 ml water. Add 8.315 ml 3 M KCl. Add KOH and adjust pH 6.8. Add MnCl2・4H2O. Add water up to 200 ml. Filter sterilize

DNS reaegnt

Materials	Methods
4.5% NaOH : 30 ml 1% DNS : 88 ml Rochelle salt : 25.5 g 10% NaOH : 2.2 ml Phenol : 1 g Water : 7.8 ml NaHCO3 : 0.69 g	Add 1% DNS 88ml and Rochelle salt 25.5 g to 4.5% NaOH 30 ml (A solution) Add Phenol 1 g and water 7.8 ml to 10% NaOH 2.2 ml (B solution) Add NaHCO3 6.9g to B solution 6.9 ml Add A solution 118ml to B solution 6.9 ml Leave 2 days Store in brown bottle