Team:PKU Beijing/Notebook/Bistable/Hao Wang

Notebook > Bistable > Hao Wang's Note

2009.8.3
13:45 Start to make LB(s)

14:16 Mini prep plasmid T7P+C1; bi-stable

15:37 Starve the colony T7P+C1; bi-stable

17:34 Start the transformation Make the A+K+ LB plate

19:21 Start the incubation

2009.8.6
13:00 Transfromation: 1-23H; T7P+C1; bi-stable

2009.8.7
9:20 Start the incubation: 1-23H; T7P+C1; bi-stable

22:19 Mini-prep plasmid: 1-23H

22:50 Prepare PCR: lacZ alpha fragment

23:25 PCR start

2009.8.8
18:38 PCR results. Extraction 1-23H-rebuild (1-23Hr)

19:34 Prepare the enzyme solution:

19:32 Start the cut

22:30 Connect 1-18C to 1-23Hr

2009.8.9
8:00 Transfromation: 1-18C+1-23Hr

22:44 Incubation 1-18C+1-23Hr

2009.8.10
8:20 Incubation Bi-stable (BL21, Kan+)

9:14 Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)

19:46 Pick the white colony: 1-18C+1-23H, incubate it

2009.8.11
8:24 Bi-stable (BL21) incubation

9:24 1-18C+1-23H, +X-gal (DMF), incubation

13:01 1-18C+1-23H, spread on the plate (Amp+)

2009.8.12
19:40 Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)

19:45 Start the incubation

19:51 spread X-gal (DMF) 50uL on the plate (Amp+)

21:19 A600=0.136

21:42 A600=0.216

22:08 A600=0.353

22:33 A600=0.467

22:40 +1mL K+ LB, ( too late, give up)

2009.8.13
13:20 Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)

13:40 spread 1-18C+1-23H 50uL on the X-gal plate (Amp+); incubate

14:04 prep the plasmid: bi-stable, and check

2009.8.14
21:58 Transformation: hi-copy T7P+C1 -> Bi-stable

2009.8.15
18:38 spread 1-18C+1-23H 50uL on the X-gal plate

19:22 incubation: low-copy Amp+ plasmid