IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/05/22

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Testing of LB-Kan Plates

 * Tested LB-Kan plates by streaking frozen glycerol stocks of DB3.1 cells on LB plate and LB-Kan plate.
 * Expect growth on LB plate and no growth on LB-Kan plate.
 * Incubated at 37 deg C in the Lagally Lab Biohazard room at 11AM.

Transformation and Calculation of Cell Competency
Example calculation: 1uL of 10pg/uL pUC19 pDNA + 100uL competent cells + 400uL LB recovery media 400uL plates are 8*10-12g giving rise to 29 or 25 colonies. 29cfu/(8*10-6μg) = 3.6*106cfu/μg

Made Chloramphenicol

 * 25mg/mL stocks; 20 aliquots of 1 mL each, therefore 20 mL
 * Dissolved 500 mg (0.5 g) of Chloramphenicol into 20 mL of 100% Ethanol
 * Aliquoted 1 mL ea. into sterile microfuge tubes, freezed at -20ºC

Extracted BioBricks from the wells

 * Resuspended the BioBricks from inside their wells with 15μL diH2O
 * Extracted the BioBricks into microfuge tubes

Transformation of Biobricks

 * Amount plated = 500uL
 * Amount of biobrick transformed = 1uL
 * amount of competent cell = 100uL of each.

Notes:
 * hot water bath was hotter than normal: 46ºC rather than 42ºC.
 * incubated for: 1 hour and 15 minutes(3:45pm to 5:00pm) @ 31ºC.
 * Tetracycline plate was left out in the light for about a hour. May cause degradation of tetracycline.
 * Incubating over the weekend (Friday night to Monday morning) at room temperature