Team:Hawaii/Notebook/2008-08-16

= Things we did today =

Reconstruction of BB-pRL1383a (cont.)

 * Grace


 * Ran digests on a 1% agarose gel
 * J33207 bands @ ~900bp and smudge ~100bp. ~100bp=VF/VR? Too much DNA loaded = band ran slower?
 * BB-pRL1383a band @ ~10kb. No band ~750bp. XbaI/PstI did not cut??
 * Extracted bands from gel
 * Ligated:
 * 1 &mu;l XbaI/PstI digested BB-pRL1383a with 3 &mu;l XbaI/PstI digested J33207
 * 1 &mu;l XbaI/PstI digested BB-pRL1383a to itself (negative control)
 * Transformed into DH5&alpha;
 * Used 5 &mu;l ligation reaction to transform
 * Plated on LB+sp100+X-gal and LB+sp100+sm50+IPTG+X-gal
 * To verify if IPTG is needed for lacZ expression

Plasmid preps

 * Grace


 * Resuspended plasmid preps in 50 &mu;l TE buffer and stored in -20C freezer (long green tray)

= Discussion =

= Quote of the Day = "She grew on him like she was a colony of E. coli, and he was room temperature Canadian beef. - Courtesy of Help.com"