Team:Penn/Protocols
From 2012.igem.org
(Difference between revisions)
Line 127: | Line 127: | ||
<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> | ||
- | </ | + | |
+ | |||
+ | <h2>Pelleting & Sonication </h2> | ||
+ | <li>5. Collect cultures by centrifugation for 30 minutes @4°C & 5000g </li> | ||
</p> | </p> |
Revision as of 05:19, 30 September 2012
Protein Purification
RSB250-A Recipe:
- 20mM Tris
- 250mM NaCl
- 30mM Imidazole
- 10% Glycerol (v/v)
- 20mM Tris
- 250mM NaCl
- 500mM Imidazole
- 10% Glycerol (v/v)
Induction
- Grow a small booster culture in LB Broth overnight (~2-10% of final culture volume) w/ appropriate antibiotics
- Innoculate full scale culture & monitor OD600 every 15-30 minutes until OD600 reaches 0.8
- At OD600=0.8, induce with appropriate concentration of inducer (e.g. 1mM final concentration of IPTG for lac promoter)
- Allow to grow overnight @ 25°C
- 5. Collect cultures by centrifugation for 30 minutes @4°C & 5000g