IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/06/30
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Removed plates from incubator at 0840 Results:
This confirms the psB1C3 backbone from last year does contain ccdB. This is useful information because we've been operating under the assumption of the presence of ccdB despite it being absent from this year's parts registry. Eric Finlay 12:21, 30 June 2010 (EDT)
Poured 12 kanamycin antibiotic plates. Last bottle of 400ml LB agar used. Need to make more LB
1. Checked the status of transformations. 5/6 plates had at least 2 colonies show up. Will leave plate 6 overnight to see if anything grows. Control plate had too many to count.
2. PCR'd 2 colonies from each plate to verify that they contain the parts desired (promoter+ RBS + fluorescence protein + terminator). Using Vf2 and Vr primers, the length should be approximately 1-1.1 kb.
3. Ran a gel of the PCR products using 110 V , 0.8% agarose, and 45 minutes run time.
4. Gel showed no bands. 100 bp ladders (flanking the PCR product lanes) also did not show up. Problem could be stain deactivation due to high temperature when pouring the gel (stain was added to the molten agarose). Will run the gel again with remaining PCR product tomorrow. Stain will be added with agarose is cooler.
Made 2 new glycerol stocks of staph (Staph obtained from liquid overnight culture prepared on June 29th. Taken out on June 30th)
Marianne and Vicki
Note: MME and MMF calculations include 80 rxns + 2 extra (for pipeting error) + H2O controls (3); original master mix in Table 1 calculations only contain 80 rxns.
Start time: 1013