Team:Wash U/Protocol

From 2009.igem.org

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(→Recipes)
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*15g Agar
*15g Agar
*up to 1 Liter H20
*up to 1 Liter H20
 +
'''ATCC medium: Van Niel's yeast agar'''
 +
*1.0 g K2HPO4
 +
*0.5 g MgSO4
 +
*10.0 g yeast extract
 +
*20.0 g agar
 +
*1.0 L tap water
 +
*Adjust pH to 7.0-7.2
'''Super Optimal Broth (SOB)'''
'''Super Optimal Broth (SOB)'''
* 2% w/v bacto-tryptone (20g)
* 2% w/v bacto-tryptone (20g)

Revision as of 20:04, 29 June 2009

Procedures

Polymerase Chain Reaction (PCR)

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Transformation

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Biobrick Assembly

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Gel Electrophoresis

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Recipes

Lysogeny Broth (LB) Media

  • 10g Tryptone
  • 5g Yeast Extract
  • 10g NaCl
  • 15g Agar
  • up to 1 Liter H20

ATCC medium: Van Niel's yeast agar

  • 1.0 g K2HPO4
  • 0.5 g MgSO4
  • 10.0 g yeast extract
  • 20.0 g agar
  • 1.0 L tap water
  • Adjust pH to 7.0-7.2

Super Optimal Broth (SOB)

  • 2% w/v bacto-tryptone (20g)
  • 0.5% w/v bacto-yeast extract (5g)
  • 8.56mM NaCl (0.5g) or 10mM NaCl (0.584g)
  • 2.5mM KCl (0.186g)
  • ddH2O to 1L

Super Optimal Broth with Catabolite repression (SOC)

  • In addition to all of the SOB components,
  • 10mM MgCl2 (0.952g)
  • 20mM glucose (3.603g)

Gel (for Electrophoresis)

  • 50 mL 1X TAE buffer
  • 0.8 g agarose
  • 2.5 microliters EtBr

Tris EDTA (TE) buffer

  • 750 mL d-H20
  • 242 g Tris base
  • 57.1 mL glacial acetic acid
  • 100 mL 0.5M EDTA (93.05 g EDTA in 500 mL d-H20, pH ~8.0)
  • fill to 1 L
  • adjust pH to 8.5


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