Team:Kyoto/Medium
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- | # Stir dry yeast and agarose with about two-thirds of water. Then, autoclave it. | + | =='''Medium'''== |
- | # Stir corn flour and glucose with the remaining water. | + | === Standard medium for drosophila === |
+ | |||
+ | # Stir dry yeast 20g and agarose 3.5~5.0g with about two-thirds of water 500mL. Then, autoclave it. | ||
+ | # Stir corn flour 45g and glucose 50g with the remaining water. | ||
# Stir 1 and 2, then autoclave it again. | # Stir 1 and 2, then autoclave it again. | ||
- | # after autoclave, add propionic acid and 10 % p-hydroxybenzoate in 70 % Eternol into it. | + | # after autoclave, add propionic acid 1.5mL and 10 % p-hydroxybenzoate in 70 % Eternol 5mL into it. |
- | [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html | + | [http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Standard medium for drosophila] |
===M9 medium=== | ===M9 medium=== |
Latest revision as of 03:14, 6 October 2011
Contents |
Medium
Standard medium for drosophila
- Stir dry yeast 20g and agarose 3.5~5.0g with about two-thirds of water 500mL. Then, autoclave it.
- Stir corn flour 45g and glucose 50g with the remaining water.
- Stir 1 and 2, then autoclave it again.
- after autoclave, add propionic acid 1.5mL and 10 % p-hydroxybenzoate in 70 % Eternol 5mL into it.
[http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Standard medium for drosophila]
M9 medium
- Stir Na2HPO4 6 g, KH2PO4 3 g, NaCl 0.5 g and NH4Cl 1 g with water 1 l.
- 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
- Stir Tryptone 20 g, Yeast extract 10 g and NaCl 10 g with water 200ml.
- If you make LB plates, add agar 10 g.
- Autoclave.
SOB medium
- Stir Tryptone 20g and Yeast extract 5g with water.
- Add 2 ml 5 M NaCl and 840 ul 3 M KCl and add water up to 1 l.
- After autoclave, add 10 ml filter sterilized 1 M MgSO4 and 1 M MgCl2.
SOC medium
- Add 2 M glucose 1 ml to 100 ml SOB.
Buffer TB
- 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
- 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