IGEM:IMPERIAL/2006/LabCalendar/2006-8-23

S01656 Testing

 * Measure OD of cultures
 * +IPTG = 1.735
 * -IPTG = 0.298 (lack of oxygen in culture, since cultured 50 mL in 50 mL Falcon tube)
 * Dilute down to OD 0.1 in 5 mL LB Kanamycin
 * Start shake at 9:25
 * Measure OD of cultures again
 * +IPTG = 0.389
 * -IPTG = 0.390
 * Dilute down to OD 0.1 in 5 mL LB (no antibiotic)
 * Start shake at 11:40
 * Measure OD of T9002
 * OD = 0.374
 * Spun down S01656 cultures, put into T9002, placed into 96 well plate
 * Start shake at 14:07

J37015 + J37015 RS Testing

 * Inoculated fresh day cultures at 9.40 am

T9002 & J37016 Testing

 * Measurement of OD in the morning:
 * T9002: 0.549
 * J37016: 0.676
 * Dilution to OD 0.1 & put in shaker at 9:45 am
 * Measurement of OD after 2h in shaker:
 * T9002: 0.359
 * J37015: 0.433
 * Dilution to OD 0.1 again - then transferring solutions with appropriate concentration of AHL respectively into 96 well plate
 * Put in shaker at 12:30 pm (set shaker to 100rev/min otherwise platees might get contaminated)
 * After 4:45 h took plates out of shaker and read fluorescence with Wallac Victor III

LoxP PCR

 * Inconclusive results - further tests tomorrow
 * The problem that we have is that the amplified bands are of a very similar size to that of the oligonucleotides, and therefore distinguishing them is proving difficult.
 * We are altering the compositions of the gels as a consequence.

Storing Bacteria

 * Following the MIT protocol S01656, T9002, J37016, J37015 & J37015RS have been stored in the -80C freezer. (They were stored with an initial OD of 0.5)

Cultures

 * Cultures set up for testing tomorrow:
 * 2 S01656 in 2ml LB KAN (-IPTG)
 * 2 S01656 in 2ml LB KAN (+ 20 IPTG)
 * 2 T9002 in 2ml LB AMP
 * 2 J37016 in 2ml LB AMP
 * 2 J37020 in 2ml LB AMP
 * 2 J37015 in 2ml LB AMP
 * 2 J37015RS in 2ml LB AMP