Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2016/02/10
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DBN012 (HA1:HPK:CFP:CFP:EM7:ZeoR:HA2) ligation results:
Going to pick 4 colonies and start liquid cultures.
Plasmid concentrations are all between 60-80 ng/µL
Digested with NotI at 37 °C for 10 minutes, then ran on the gel at 100 V for 60 minutes. Expect a single band at 5875 bp (no NotI cut site in HA1 part).
Colonies 2 and 3 look good, 1 is hard to tell but looks okay, 4 is not good. Going to proceed with sequencing colonies 2 and 3, using both biobrick forward and reverse primers.