Lab Manual
biological reagents
Cell Free Systems
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misc methods

Induction Protocol for TAC Overexpression Vectors

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Protocol for small scale induction to test expression levels

This protocol is used to do a time course or other optimizations prior to scale up to optimize overexpression.

For example a sample time course in minutes might be:
t= 0,30,60,90,120,... post induction

  1. Transform cells and pick a fresh colony into 5 mls LB amp Grow cells to saturation overnight Dilute cells 1:10 (200 microliters in 2 ml) with LB amp and grow further 180min (3 hrs) at 37ºC. Induce with 1mM final IPTG (100mM stock 23.8 mg/ml) Grow 2 hours more at 37ºC.  Boil prep (see small scale protocol )
  2. Western Blot.