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_I732006 | 09Kit 1 | 23H | PSB1AK3 | LacZ 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:
| 5ul | plasmid: 1-23Hr
|
| 1ul | X-bal
|
| 1ul | Pst1
|
| 2ul | bufferM
|
| 11ul | ddH2O
|
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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