Team:PKU Beijing/Notebook/Bistable/Hao Wang

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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

BBa_I73200609Kit 123HPSB1AK3LacZ Fragment

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:

5ulplasmid: 1-23Hr
1ulX-bal
1ulPst1
2ulbufferM
11ulddH2O

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

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