Team:Hawaii/Notebook/2008-08- 8

= Things we did today =

Restreaked nir+rbs and I14032+rbs constructs

 * Grace

Plasmid prep (finished up)

 * Grace


 * Resuspended plasmid preps in 50 &mu;l TE buffer
 * Determined DNA concentrations of plasmids

Prep for sequencing

 * Grace
 * 25 &mu;l PCR reactions of nir+rbs, I14032+rbs, slr1, slr2, BB-pRL1383a
 * Colony PCRs seem to indicate success
 * 25 &mu;l PCR reactions of GFP+tt, GFPf+tt, J33207+tt
 * Colony PCR indicates no ligation. Picked new colony, sequence to confirm failure.
 * PCR of nir, B0015, B0030, B0034 for resequencing (bad reads last time)
 * Gel purified all PCR rxns (we still have a problem with contaminant DNA/shadow bands) and desired bands were extracted from gel
 * 2% agarose gel ran at 60v for 2 hours
 * Determined DNA concentrations via nanodrop spectrometer
 * Prepared samples and sent to CORE Hawaii for sequencing

Construct p+r+g and p+r+s

 * Krystle


 * Restriction Digest
 * nir+B0030, I14032+B0030, J33207 digested with SpeI and PstI
 * gfp, gfpfusion, and B0015 digested with XbaI and PstI
 * Gel Purified restriction digest
 * 2% agarose gel ran at 60 volts for 1.5 hours
 * 40ul of the total restriction digest loaded into each well
 * gfpfusion cut out from gel

= Discussion =

= Quote of the Day = "History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson"