Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2016/01/29

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Overview

QuikChange of EM7:ZeoR_v0120 and MV8, continued...

Treated the EM7:ZeoR_v0120, MV8, and half of pWhitescript with 1 µL DpnI restriction enzyme. Left half of pWhitescript untreated as a control to see change in transformation efficiency.

Set up four LB-Amp plates, transformed using quick transformation protocol. The two control plates (pWhitescript) were first coated with 100 µL IPTG (10 mM) and 40 µL X-Gal (5% w/v) each, to induce expression of β-galactosidase and produce blue colonies for successful transformation. Will be counting # of blue and white colonies on each of the control plates as a measure of the efficiency of the QuikChange protocol.

Plates were incubated at 37 °C for 4 hours, then left on the bench at room temperature. Will be transferred to the fridge by Daniel tomorrow afternoon.