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Team:Cambridge/Protocols/IPTGInduction
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1) Pick cultures and grow in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube overnight at 37<sup>0</sup>C at 160RPM. | 1) Pick cultures and grow in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube overnight at 37<sup>0</sup>C at 160RPM. | ||
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2)Dilute 1:100 and grow for 3-4 hours in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube | 2)Dilute 1:100 and grow for 3-4 hours in 2ml LB/AMP (100ug/ml) in a 15ml snap cap tube | ||
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3)Spread 250uL and 25uL onto you IPTG plates. Also spread some on normal amp plates as a control. Allow to grow in incubator. | 3)Spread 250uL and 25uL onto you IPTG plates. Also spread some on normal amp plates as a control. Allow to grow in incubator. | ||
Revision as of 12:04, 22 August 2012
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.
