Team:Penn/Protocols

From 2012.igem.org

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<li> 10% Glycerol (v/v) </li>
<li> 10% Glycerol (v/v) </li>
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</ul>
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<h2>Induction</h2>
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1. Grow a small booster culture in LB Broth overnight (~2-10% of final culture volume) w/ appropriate antibiotics
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2. Innoculate full scale culture & monitor OD600 every 15-30 minutes until OD600 reaches 0.8
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3. At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter)
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4. Allow to grow overnight @ 25°C

Revision as of 05:15, 30 September 2012

Penn 2012 iGEM Wiki

Protein Purification

RSB250-A Recipe:

  • 20mM Tris
  • 250mM NaCl
  • 30mM Imidazole
  • 10% Glycerol (v/v)
RSB250-B Recipe
  • 20mM Tris
  • 250mM NaCl
  • 500mM Imidazole
  • 10% Glycerol (v/v)

Induction

1. Grow a small booster culture in LB Broth overnight (~2-10% of final culture volume) w/ appropriate antibiotics 2. Innoculate full scale culture & monitor OD600 every 15-30 minutes until OD600 reaches 0.8 3. At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter) 4. Allow to grow overnight @ 25°C

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