2009.7.23
10:30
Double-enzyme digestion: 1-23L, 1-2M+AraC
16:50
Gel purification
19:00
Ligation:
Insert: 1-2M+AraC
Vector: 1-23L
23:00
Double-enzyme digestion
2009.7.24
Gel purification
2009.7.27
19:00
Ligation:
Insert: 1-2M+AraC+1-23L
Vector: Pcat
23:00
Transformation
2009.7.28
11:00
PCR: Pcat+1-2M+AraC+1-23L
23:00
Mini-prep and store strains: 1, 2, 4, 5, 6, 7
BioSpec(×50 μg/mL):
| 1 | 2.5823
|
| 2 | 2.5939
|
| 4 | 3.0660
|
| 5 | 2.5124
|
| 6 | 2.2350
|
| 7 | 2.2235
|
00:30
Reverse PCR: Pcat+1-2M+AraC+1-23L
2009.7.29
12:30
Gel purification
BioSpec(×50 μg/mL): 0.1964
14:00
Double-enzyme digestion: Reverse PCR fragment and plasmid of Pcat+1-2M+AraC+1-23L
20:30
Gel purification
22:00
Double-enzyme digestion: 1-1F (Front vector)
2009.7.30
13:30
Gel purification: 1-1F
15:00
Ligation:
Insert: Pcat+1-2M+AraC+1-23L
Vector:1-23L
21:00
Transformation: ligation product
2009.7.31
10:30
PCR
16:00
PCR
23:00
Mini-prep
2009.8.1
12:00
Transformation
2009.8.2
11:30
Pouring plates: LB+Amp ×30, LB+Kan ×50
15:30
Mini-prep: pla1~5, pcr1~4
Gel electrophoresis
17:00
Double-enzyme digestion (front insert)
Plasmid: pla1, pla2, pcr1, pcr3
22:30
Gel purification (pcr1, pla1, pla2)
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