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From 2012.igem.org

week 15: 6 august - 12 august

Office work

Lab work

New electrocompetent DH5 Alfa and Mach1 E. coli cells were prepared. Transformation efficiency was determined: DH5 Alfa: 10^8 Mach1: 10^9

TuYV

• Bricking:

Last week's minipreps were digested (EcoRI, PstI), the inserts ligated into the chloramphenicol backbone from BBa_J04450 and transformed into our electrocompetent DH5 Alfa E. coli. A PCR check gave negative results, but miniprep was performed anyway.

• Expression:

Last week's minipreps were digested (XbaI, Spe1), the inserts ligated into the promotor&RBS-containing backbone from BBa_J04500 and transformed into our electrocompetent BL21 E. coli. Results were unsatisfying at first, so the entire procedure was done again on saturday.

Written by: Wouter

PLRV

Four different PLRV CP's were transformed into E.coli DH5α electrocompetent cells. After culturing the cells, we miniprepped the plasmids. Restriction check with XbaI and SpeI enzymes confirms the presence of the PLRV CP inside the plasmid.

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