Team:Wash U/Protocol

From 2009.igem.org

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'''Polymerase Chain Reaction (PCR)'''
'''Polymerase Chain Reaction (PCR)'''
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'''Transformation'''
'''Transformation'''
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<html><a href="http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.jpg"><img align="right" height=200px width=200px align="right" src="https://static.igem.org/mediawiki/2009/c/c5/BioBrick_Assembly_Manual.jpg"></a></html>
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<html><a href="http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.jpg"><img align="right" height=225px width=250px align="right" src="https://static.igem.org/mediawiki/2009/c/c5/BioBrick_Assembly_Manual.jpg"></a></html>
'''Biobrick Assembly'''<br>
'''Biobrick Assembly'''<br>
:To view the full Biobrick Assembly manual, click [http://ginkgobioworks.com/support/ here].
:To view the full Biobrick Assembly manual, click [http://ginkgobioworks.com/support/ here].
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'''Gel Electrophoresis'''
'''Gel Electrophoresis'''
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Revision as of 15:36, 30 June 2009

Procedures

Polymerase Chain Reaction (PCR)

text



Transformation

text



Biobrick Assembly

To view the full Biobrick Assembly manual, click here.







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)

TAE Buffer

  • 242 g Tris base (2-amino-2-hydroxymethyl-propane-1,3-diol) (= 2 mole)
  • 57.1 ml glacial acetic acid (= 100% acetic acid) (57.19 ml = 1 mole)
  • 100 ml 0.5 M Na2 EDTA (pH 8.0)
  • ddH2O to 1L

TBE Buffer

  • 53 g of Tris base (CAS# 37186)
  • 27.5 g of boric acid (CAS# 11280)
  • 20 ml of 0.5 M EDTA (CAS# 60004) (pH 8.0)

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

Gel (for Electrophoresis)

  • 50 mL 1X TAE buffer
  • 0.8 g agarose
  • 2.5 microliters EtBr
  • Note: Jacob suggested adding 1.0 microliter EtBr to gel and 3.0 microliters to TAE buffer in rig.


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