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Team:Cambridge/Protocols/IPTGInduction
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This protocol is an adaptation of this one: http://www.genetics.wustl.edu/tslab/?page_id=107 | 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 37<sup>0</sup>C 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. | ||
{{Template:Team:Cambridge/CAM_2012_TEMPLATE_FOOT}} | {{Template:Team:Cambridge/CAM_2012_TEMPLATE_FOOT}} | ||
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.
