Purification of GST Fusion Proteins: Difference between revisions
Created page with '==Bacteria Production and Induction== #Express and induce protein in culture under appropriate conditions (normally do the following) #grow an overnight culture in ~25 mL LB/Amp ...' |
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#add 5 mL culture to 1L TB/Amp and grow at 37C | #add 5 mL culture to 1L TB/Amp and grow at 37C | ||
#grow to OD600 of 0.6-1.0 and induce with 10 uM IPTG (optimize for each protein) | #grow to OD600 of 0.6-1.0 and induce with 10 uM IPTG (optimize for each protein) | ||
#let grow O/N at <25C (optimize induction time/temp for each protein). | #let grow O/N at <25C (optimize induction time/temp for each protein, see [[Induction Conditions]]). | ||
#centrifuge cells 10 min at 9000 RPM to pellet bacteria freeze if stopping at this point | #centrifuge cells 10 min at 9000 RPM to pellet bacteria freeze if stopping at this point | ||
==Lysis and Purification== | ==Lysis and Purification== | ||