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Transforming bacteria: Difference between revisions
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→Transformation Wed 11 sept 2013
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* Incubated 50ul of competent DH5α from 2013-4-11 with 2ul PUC19 for 30 mins on ice (also control with no PUC19 added). Then 42C for 45 seconds. 2 mins back on ice. Added 500ul LB. Incubated at 37C for 1 hr. Plated on amp plates and incubated at 37C overnight. | * Incubated 50ul of competent DH5α from 2013-4-11 with 2ul PUC19 for 30 mins on ice (also control with no PUC19 added). Then 42C for 45 seconds. 2 mins back on ice. Added 500ul LB. Incubated at 37C for 1 hr. Plated on amp plates and incubated at 37C overnight. | ||
* After ~ 15 hours there was a small cluster of colonies. (Incubator had said it was at 42C). Control was empty. Left at RT for another 22 hours. | * After ~ 15 hours there was a small cluster of colonies. (Incubator had said it was at 42C). Control was empty. Left at RT for another 22 hours. | ||
* > 100 colonies on the plate. Still nothing on control. Replated a single colony onto another amp plate and incubated at RT. | * > 100 colonies on the plate. Put it in the fridge. Still nothing on control. Replated a single colony onto another amp plate and incubated at RT. | ||
