Team:Brown-Stanford/Lab/Protocols/BG11

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<html><h1>Protocol Code: M1</h1></html>
<html><h1>Protocol Code: M1</h1></html>

Revision as of 17:38, 27 September 2011

Brown-Stanford
iGEM
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Protocol Code: M1

BG-11

(Taken from Meeks Lab at University of California, Davis)

Stock solutions for BG-11

Stock 1

  • 0.1g/liter Na2MG EDTA
  • 0.6g/liter Ferric ammonium citrate
  • 0.6g/liter Citric acid•1H2O
  • 3.6g/liter CaCl2•2H2O

Filter sterilize into a sterile bottle or autoclave.

Stock 2

  • 7.5g/liter MgSO4•7H2O

Filter sterilize into a sterile bottle or autoclave.

Stock 3

  • 4.0g/liter K2HPO4•3H2O or 3.05g/liter K2HPO4

Filter sterilize into a sterile bottle or autoclave.

Stock 5 (Microelements)

Stock 5 (Microelements):

  • 2.86g/liter H3BO3
  • 1.81g/liter MnCl2•4H2O
  • 0.222g/liter ZnSO4•7H2O
  • 0.079g/liter CuSO4•5H2O
  • 0.050g/liter COCl2•6H2O
  • 0.391g/liter NaMoO4•2H2O or 0.018g/liter MoO4 (85%)

Basic BG11 medium

Combine the following stock solutions (for 1 liter):

  • 10 mL Stock 1
  • 10 mL Stock 2
  • 10 mL Stock 3
  • 0.02 g Na2CO3
  • 1.0 mL Stock 5
  • 1.5 g NaNO3

Combine stocks and adjust pH to 7.5 (use 1.0N HCl). Aliquot into flasks (50 mL/125 mL flask) with cotton stoppers on top and autoclave. After autoclaving and cooling the pH is about 7.1. For solid media add 1% noble agar. For BG-11, don't add NaNO3.

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