IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/09/27

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Biofilm Protocol Day 4

 * 15 minute Crystal Violet Stain on plates EF09251020-1 to EF09251030-3

DspB Track

 * O/N culture for sonication taken out at 12pm; placed in 4C
 * Results: all tubes cloudy; J23100 is slightly red, rest is yellow colour
 * Made 1/100 dilutions for pre-dspB & 14 (Pcon + RBS) with C1 antibiotic
 * Put in 37C incubator at 12:15pm


 * Results: Lid closed too tight --> dspB = 0.081; 14 = 0.477
 * Final OD: pre-dspB = 1.15; 14 = 0.91
 * Centrifuge down into pellet
 * 1 mL into each microcentrifuge each time at 6000rcf for 5min each time (all 5mL)

Rafael's Method

 * Mix rxn 1 with rxn 2 in rxn1 tube
 * Run in PCR machine:
 * 3 cycles:
 * 65C @ 10 min
 * 37C @ 20 min


 * Start: 8pm
 * End: 9:33pm

Ligation- Rafael's Method

 * Tubes: Raf
 * Content: backbone XP + insert (1) DspB
 * Incubate at room temperature for 20min
 * Store in -20C


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