Team:Hawaii/Notebook/2008-07-15

= Things we did today =

Oligonucleotide experiment, continued

 * Grace


 * Ran overnight RE digests of pRL1383a and BBa_C0012 on EtBr stained 2% agarose gel at 95V for 1 hour
 * BBa_C0012 vector okay
 * pRL1383a vector band ~9kb; no band observed at ~350bp. RE did not cut?
 * Gel purified pRl1383a, BBa_C0012, GFP fusion, and Biobrick segments
 * Used nanodrop spectrometer to determine DNA concentration (below)
 * Ligated 7 &mu;l Biobrick segment with 0.5 &mu;l pRL1383a and 7 &mu;l GFP fusion with 2 &mu;l BBa_C0012 vector in 20 &mu;l ligation reactions
 * Transformed DH5&alpha; using 10 &mu;l ligation reactions
 * Plated GFP/C0012 vector on LB + amp100
 * Plated BB segment/pRL1383a on LB + sp100
 * Counted colonies from yesterday's pnir, slr2016, pilA assembly transformation (see experiment)
 * Colony PCR'd colonies to verify presence of desired products

Transformation
Margaret


 * Transformation of DB3.1 competent cells pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-) no plasmid, because transformation from 7/14/08 yielded only 1 colony from pSB3K3, nothing from pSB1A2 & pSB1AK3.

Plasmid prep
Krystle


 * Resuspended yesterday's plasmid prep in 100 &mu;l TE and stored in -20C
 * Ran plasmid preps on gel to verify
 * Grew up pnir, slr2016, and pilA transformants in LB in preparation for plasmid prep tomorrow

Restriction Digest
Krystle


 * RE digested BBa_B0034, BBa_B0024, and BBa_E0040 with XbaI overnight

= Discussion =

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