Team:PKU Beijing/Notebook/Bistable/Hao Wang

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(Difference between revisions)
(New page: {{PKU_Beijing/Header}} {{PKU_Beijing/Sidebar_Notebook}} {{PKU_Beijing/Header2}} ==='''2009.8.3'''=== '''13:45''' Start to make LB(s) '''14:16''' Mini prep plasmid T7P+C1; bi-stable '''1...)
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==='''2009.8.3'''===
==='''2009.8.3'''===
-
'''13:45'''
+
'''13:45'''<br>
Start to make LB(s)
Start to make LB(s)
-
'''14:16'''
+
'''14:16'''<br>
Mini prep plasmid T7P+C1; bi-stable
Mini prep plasmid T7P+C1; bi-stable
-
'''15:37'''
+
'''15:37'''<br>
Starve the colony T7P+C1; bi-stable
Starve the colony T7P+C1; bi-stable
-
'''17:34'''
+
'''17:34'''<br>
-
Start the transformation
+
Start the transformation<br>
Make the A+K+ LB plate
Make the A+K+ LB plate
-
'''19:21'''
+
'''19:21'''<br>
Start the incubation
Start the incubation
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|}
|}
-
'''13:00'''
+
'''13:00'''<br>
Transfromation: 1-23H; T7P+C1; bi-stable
Transfromation: 1-23H; T7P+C1; bi-stable
==='''2009.8.7'''===
==='''2009.8.7'''===
-
'''9:20'''
+
'''9:20'''<br>
Start the incubation: 1-23H; T7P+C1; bi-stable
Start the incubation: 1-23H; T7P+C1; bi-stable
-
'''22:19'''
+
'''22:19'''<br>
Mini-prep plasmid: 1-23H
Mini-prep plasmid: 1-23H
-
'''22:50'''
+
'''22:50'''<br>
Prepare PCR: lacZ alpha fragment
Prepare PCR: lacZ alpha fragment
-
'''23:25'''
+
'''23:25'''<br>
PCR start
PCR start
==='''2009.8.8'''===
==='''2009.8.8'''===
-
'''18:38'''
+
'''18:38'''<br>
PCR results. Extraction 1-23H-rebuild (1-23Hr)
PCR results. Extraction 1-23H-rebuild (1-23Hr)
-
'''19:34'''
+
'''19:34'''<br>
-
Prepare the enzyme solution:
+
Prepare the enzyme solution:<br>
{|cellpadding=5
{|cellpadding=5
|5ul||plasmid: 1-23Hr
|5ul||plasmid: 1-23Hr
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|}
|}
-
'''19:32'''
+
'''19:32'''<br>
Start the cut
Start the cut
-
'''22:30'''
+
'''22:30'''<br>
Connect 1-18C to 1-23Hr
Connect 1-18C to 1-23Hr
==='''2009.8.9'''===
==='''2009.8.9'''===
-
'''8:00'''
+
'''8:00'''<br>
Transfromation: 1-18C+1-23Hr
Transfromation: 1-18C+1-23Hr
-
'''22:44'''
+
'''22:44'''<br>
Incubation 1-18C+1-23Hr
Incubation 1-18C+1-23Hr
==='''2009.8.10'''===
==='''2009.8.10'''===
-
'''8:20'''
+
'''8:20'''<br>
Incubation Bi-stable (BL21, Kan+)
Incubation Bi-stable (BL21, Kan+)
-
'''9:14'''
+
'''9:14'''<br>
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
-
'''19:46'''
+
'''19:46'''<br>
Pick the white colony: 1-18C+1-23H, incubate it
Pick the white colony: 1-18C+1-23H, incubate it
==='''2009.8.11'''===
==='''2009.8.11'''===
-
'''8:24'''
+
'''8:24'''<br>
Bi-stable (BL21) incubation
Bi-stable (BL21) incubation
-
'''9:24'''
+
'''9:24'''<br>
1-18C+1-23H, +X-gal (DMF), incubation
1-18C+1-23H, +X-gal (DMF), incubation
-
'''13:01'''
+
'''13:01'''<br>
1-18C+1-23H, spread on the plate (Amp+)
1-18C+1-23H, spread on the plate (Amp+)
==='''2009.8.12'''===
==='''2009.8.12'''===
-
'''19:40'''
+
'''19:40'''<br>
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
-
'''19:45'''
+
'''19:45'''<br>
Start the incubation
Start the incubation
-
'''19:51'''
+
'''19:51'''<br>
spread X-gal (DMF) 50uL on the plate (Amp+)
spread X-gal (DMF) 50uL on the plate (Amp+)
-
'''21:19'''
+
'''21:19'''<br>
A600=0.136
A600=0.136
-
'''21:42'''
+
'''21:42'''<br>
A600=0.216
A600=0.216
-
'''22:08'''
+
'''22:08'''<br>
A600=0.353
A600=0.353
-
'''22:33'''
+
'''22:33'''<br>
A600=0.467
A600=0.467
-
'''22:40'''
+
'''22:40'''<br>
+1mL K+ LB, ( too late, give up)
+1mL K+ LB, ( too late, give up)
==='''2009.8.13'''===
==='''2009.8.13'''===
-
'''13:20'''
+
'''13:20'''<br>
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
Prepare to make the sensitive cell: Bi-stable (BL21, Kan+)
-
'''13:40'''
+
'''13:40'''<br>
spread 1-18C+1-23H 50uL on the X-gal plate (Amp+); incubate
spread 1-18C+1-23H 50uL on the X-gal plate (Amp+); incubate
-
'''14:04'''
+
'''14:04'''<br>
prep the plasmid: bi-stable, and check
prep the plasmid: bi-stable, and check
==='''2009.8.14'''===
==='''2009.8.14'''===
-
'''21:58'''
+
'''21:58'''<br>
Transformation: hi-copy T7P+C1 -> Bi-stable
Transformation: hi-copy T7P+C1 -> Bi-stable
==='''2009.8.15'''===
==='''2009.8.15'''===
-
'''18:38'''
+
'''18:38'''<br>
spread 1-18C+1-23H 50uL on the X-gal plate
spread 1-18C+1-23H 50uL on the X-gal plate
-
'''19:22'''
+
'''19:22'''<br>
incubation: low-copy Amp+ plasmid
incubation: low-copy Amp+ plasmid
{{PKU_Beijing/Foot}}
{{PKU_Beijing/Foot}}
__NOTOC__
__NOTOC__

Revision as of 10:02, 2 October 2009
 

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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