Team:Kyoto/Protocol

From 2011.igem.org

(Difference between revisions)
(Protocol)
(SOB medium)
Line 60: Line 60:
* Tryptone: 2 g
* Tryptone: 2 g
* Yeast extract : 0.5 g
* Yeast extract : 0.5 g
-
* 5M NaCl : 0.5 g
+
* 5M NaCl : 200 ul
-
* 3M KCl : 1 g
+
* 3M KCl : 84 ul
* 10mM MgSO4 : 1 ml
* 10mM MgSO4 : 1 ml
* 10mM MgCl2 : 1 ml
* 10mM MgCl2 : 1 ml

Revision as of 05:19, 23 September 2011

Contents

Protocol

Medium for drosophila

[http://www.biol.se.tmu.ac.jp/fly/www/standard-medium.html Medium for drosophila]
Materials Methods
  • water : 500mL
  • dry yeast : 20g
  • corn flour : 45g
  • glucose : 50g
  • agarose : 3.5~5g
  • propionic acid : 1.5mL
  • 10% p-hydroxybenzoate in 70% Eternol : 5g
  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.■


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
  1. Stir Na2HPO4, KH2PO4, NaCl and NH4Cl with water.
  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 10ml filter sterilized 20% casamino acid.


SOB medium

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