Team:Hawaii/Notebook/2008-08- 1

= Things we did today =

Restriction Digest

 * Grace


 * Sequential digest of B0015 with XbaI and EcoRI

Checked transformants from yesterday's ligations

 * Grace

Transformation of DB3.1 with nir and gfp constructs

PCR

 * Grace & Krystle


 * Colony PCR'd all transformants from yesterday's ligations and slr1, slr2, nir, pilA, GFPf
 * Annealed at 58C, elongated for 90 sec.
 * PCR of E0240 and I14032 from filter paper
 * Annealed at 58C, elongated for 90 sec.
 * Ran on 2.5% gel
 * Contaminant at 250bp
 * pRL1383a = 5 bands (~1.2kb, ~0.9kb, ~0.35kb, ~0.3kb, ~0.25kb); desired band = 958bp
 * nir+rbs = 2 bands (~0.275kb, ~0.25kb); desired band = 353bp
 * nir was not inserted
 * E/X sites not compatible; ligase ligated blunt ends created by RE digest leftovers?
 * GFPf+tt = 2 bands (~0.32kb, ~0.25kb); desired band = 1081bp
 * GFPf not inserted
 * Extracted E0240, slr1, slr2, GFP, pilA, I14032 from gel (correct sizes)
 * GFPf = ~1150bp (too big, what's going on? we're consistently getting GFPf this big)
 * nir = ~1100bp (too big)
 * Since sequencing returned correct sequences for GFPf and nir, regrow E. coli from colony used for sequencing and see if desired/expected results can be obtained or if results are the same

Inoculated LB+amp100

 * Grace


 * GFP
 * nir

Made LB media

 * Krystle

= Discussion =

= Quote of the Day = "Logically, it makes sense, but we're not always very logical people so it's still a weird numbering system - KS, GK in reference to the BioBrick part numbers"