Embarssing problem

[Ozan Aygun]

Hi there,

I am routinely using the same vector.
I reccomend you to use IPTG to a final of 1mM.
Moreover, start inducing at OD600:  0.2.  Leave
temperature at 37 C, whic is OK to see only GST
expression. Worth to keep some Glucose before
induction.

May worth to replace or re-prepare your IPTG stock as
well. (In Bl21-DE-RIL strain there is an internal
reporter plasmid that shows you up on the coomassie
stained gel when the IPTG induction indeed works).

If nothing works, then you may better to sequence the
promoter and operator sequences of your backbone
vector to check any modification from the original
one.

Good luck!

Ozan



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