High Stabilitiy Plasmid Development
Abstract
- P1004 Band Isolation corroboration gel, and ligation concentration estimation gel.
- Ligation of P1004 with B0014
- Ligation of repC-PCR with B0015
Electrophoresis gel
| 1.
|
Ladder 250bp
|
| 2.
|
EMPTY
|
| 3.
|
DD P1004 in pSB1A1 E,S
|
| 4.
|
Band isolated P1004
|
| 5.
|
DD KnR E,S
|
| 6.
|
DD B0014 in pSB1AK3 E,X
|
| 7.
|
DD B1007 in pSB1AK3 E,X
|
| 8.
|
DD repC-PCR E,S
|
| 9.
|
DD B0015 in pSB1AK3 st.2 E,X
|
| 10.
|
pSB1C3 digestion with XhoI (provided by Fibo)
|
Ligations
P1004 and B0014 in pSB1AK3
| Reaction
|
7:7
|
9:5
|
| P1004
|
7μL
|
9μL
|
| B0014
|
7μL
|
5μL
|
| T4 DNA ligase
|
0.5μL
|
0.5μL
|
| 5x ligase buffer
|
4μL
|
4μL
|
| H2O mQ
|
1.5μL
|
1.5μL
|
| Total
|
20μL
|
20μL
|
repC-PCR and B0015 in pSB1AK3
Both DNA solutions were too concentrated, this is why I used more water here.
| Reaction
|
6:4
|
7:4
|
| repC-PCR
|
6μL
|
7μL
|
| B0015
|
4μL
|
4μL
|
| T4 DNA ligase
|
0.5μL
|
0.5μL
|
| 5x ligase buffer
|
4μL
|
4μL
|
| H2O mQ
|
5.5μL
|
4.5μL
|
| Total
|
20μL
|
20μL
|
|
Project name
Main project page
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