Team:Penn/Protocols

From 2012.igem.org

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<li> At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter) </li>
<li> At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter) </li>
<li> Allow to grow overnight @ 25°C </li>
<li> Allow to grow overnight @ 25°C </li>
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<h2>Pelleting & Sonication </h2>
<h2>Pelleting & Sonication </h2>
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<li>5. Collect cultures by centrifugation for 30 minutes @4°C & 5000g </li>
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<li>Collect cultures by centrifugation for 30 minutes @4°C & 5000g </li>
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Revision as of 05:20, 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

Pelleting & Sonication

  • Collect cultures by centrifugation for 30 minutes @4°C & 5000g

    • Retrieved from "http://2012.igem.org/Team:Penn/Protocols"