Zulu Time All.
14-6-17
Stock Concentrations:
EBFP2: 1118 ng/µL
MAV: unknown.
1856: Diluted DNA samples for both x100 before transformation
4 Plates, LO HI for both.
2144: Culture plates incubated overnight at 30°C for 16-18 hours.
14-6-18
14-6-19
Concentrations:
EBFP2: 12 ng/µL
MAV: 4 ng/µL
1715: Culture plates inseerted into 30°C incubator.
14-6-20
1105: Low transformation rate. Suspicions fall on faulty plates. Plating to be redone later with existing samples and streaking.
14-6-23
Attempted to isolate monoclonal colony from MAV1212 lawn.
Inserted into Amp+ LB medium.
Left on incubator shaker at 2114.
Re-plated transformation suspension on Amp+ plates.
Streaked colonies from lawn.
Left on 30°C incubator at 2120.
14-6-24
Medium Clear.
Growth failed.
Current MAV1212 vectors abandoned due to overactive ccdB gene killing off cells.
Awaiting arrival of new vectors with better survivability.
Transformed onto Amp+ plates.
Left on 30°C incubator at 2135.
14-6-25
hEF1a ENTR vector transformed into competent cells and plated onto Kan+ plates.Plates incubated at 30°C incubator from 2120.
14-6-26
Culture miniprep-edConcentration:
Colonies picked into LB-Kan medium. Placed into shaker at
14-6-27
Golden Gate miR sensing sites onto MAV1212-EBFP2, replacing LacZ.
Thermo-cycler started 2220
14-6-30
Product transformed into competent cells. New plates made with Amp and X-gal.Incubation started at 2020.