IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/07/05

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BioFilm Track
from glycerol stocks
 * Grew 2 sub-cultres in 10ml of staph + B2 (3ml) for OD curve and staph competent cell. Cell obtained
 * Placed in AMBL Rotator (32C, 180rpm)
 * Start: 3:30pm (July 05,10)
 * End: 9:30am ( July 06, 10)

Misc

 * Poured Amp plates (18 plates)
 * Placed 2 ON E.coli competent cells in AMBEl rotator (32C, 180rpm)
 * Autoclaved 500ml LB Broth media
 * Autoclaved 600ml B2 media

Colony PCR

 * Protocol: Used 'colony PCR' in common protocols
 * Colony PCR for individual colonies:
 * 11T,12T
 * 5 colonies from each plate; therefore, 10 PCRs in total


 * For each respective tube, picked colonies numbered 3,5,7,9,11 in order and added to each microcentrifuge tube in ascending order. Ex. Colony #3 from plate 11T into 11T1.

PCR Cycles:
 * 95C @ 15min
 * 26 cycles:
 * 94C @ 30 sec
 * 56C @ 30 sec
 * 68C @ 2 min
 * 68C @ 20 min


 * 10C @ hold


 * Start PCR: 1351
 * End PCR: 1603

Gel verification

 * Protocol: gel verification protocol in iGEM training manual
 * Machine conditions: 110V, 40 minutes, 0.5X TBE buffer
 * DNA: 10uL, loading dye: 2uL

Gel results:

Overnight culture
Vicki Ma 18:34, 6 July 2010 (EDT)
 * Protocol: From iGEM common protocols
 * Colonies: 8T3, 6T10, 6T11, 6T14
 * Cultures (tubes): 8T3, 6T10, 6T11, 6T14, control (LB broth+chlor)
 * Put cultures in 37C turntable in AMBL lab @ 1800