Team:Cambridge/Protocols/IPTGInduction

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IPTG Induction of Ratiometrica in E. coli

This protocol is an adaptation of this one: http://www.genetics.wustl.edu/tslab/?page_id=107 and is used to test the induction of the pSPANK promoter with IPTG in E.coli

Preparation

Prepare LB/AMP/IPTG plates with 100ug/ml of Ampicillin and a final concentration of 0.5mM. Also have a couple of normal amp plates ready.

Protocol

1) Pick cultures and grow in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube overnight at 37⁰C at 160RPM. 2)Dilute 1:100 and grow for 3-4 hours in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube 3)Spread 250uL and 25uL onto you IPTG plates. Also spread some on normal amp plates as a control. Allow to grow in incubator.