May 29, 2014
Struck out fresh E. coli S17-1 onto LB only agar. Incubated at 37C overnight.
Brothed isolated colony of pBE110 (in T1R E. coli) into 10 mL of LBTet15. Incubated at 37C, shaking at 225 rpm.
May 30, 2014
Performed plasmid isolation of pBE110 using Zyppy kit. Purified plasmid DNA using Zyppy DNA clean + concentrator kit.
Made electrocompetent S17-1 E. coli cells from the plate made yesterday. Performed procedure on May 21, 2014 to make electrocompetent cells.
Electroporate transformed pBE110 into the electrocompetent S17-1 cells. Used pUC19 as control. Immediately after electroporating, added 1 mL of warmed SOC media and incubated at 37C for 1 hour. Plated pBE110 onto LBTet15Tm200 and pUC19 onto LBAmp100. Incubated at 37C overnight. Will check if have any colonies tomorrow.
//EWW
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