Team:Hawaii/Notebook/2008-07-14

= Things we did today =

PCR

 * Grace


 * 25 &mu;l PCR reactions for GFP site directed mutagenesis and extraction of Biobrick pertinent sites from BBa_B0034

Made stocks

 * Grace


 * Made two 500mL batches of SOB
 * Made 20 LB + amp100 plates

Transformed DH5&alpha; with ligations of a Biobrick vector with pnir, slr2016-1, slr2016-2, pilA, or C0012

 * Grace


 * Transformed using 5 &mu;l of 1:1 and 1:10 ligation dilutions
 * Transformed using 1 &mu;l vector only (negative control)
 * Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM

RE digested PCR products and plasmids

 * Grace


 * XbaI and PstI: GFP fusion brick and BBa_C0012
 * Added XbaI and PstI separately
 * Ran 25 &mu;l of GFP and 20 &mu;l of BBa_C0012 restriction digest reactions in gel
 * HindIII and BamHI: Biobrick segment and pRL1383a
 * Ran 25 &mu;l of Biobrick segment (half of total rxn) and 20 &mu;l of pRL1383a restriction digest reactions in gel
 * pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid

Transformed DB3.1

 * Margaret


 * Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing. The positive and negative control verify that the DB3.1 competent cells made on 7/11/08 can take up DNA. The results of this test can be seen here.

= Discussion =

= Quote of the Day = ""