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EPF-Lausanne/26 August 2009
From 2009.igem.org
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===Cloning=== | ===Cloning=== | ||
| + | We aimed to do the following cloning : | ||
| + | - LacI-RBS2 -> E0240 | ||
| + | - BB5 -> RO2 | ||
| + | |||
| + | - Klenow fragment -> I13507 | ||
===PCR=== | ===PCR=== | ||
Revision as of 09:22, 4 September 2009
Contents |
Wet Lab
Culture
The following bacteria have been cultivated and analyzed by the fluorescent microscope in Sebastian's lab : 3x RO2 (#4,5,10), 4x RO2-BB(#4-1/#5-1/#8-5/#10-6), 3x RO2 (#4,5,10).
Colony PCR
The result of the double transformation of yesterday gave the following number of clones :
RO2#10 - BB6 : ~ 30
RO2#8 - BB5 : ~ 30
RO2#5 - BB3 : a lot
RO2#4 - BB1 : a lot
There has been a double antibiotic resistance so there might be a good chance these bacteria have included both plasmids. To be sure of it, we did a colony PCR. We took 10 clones from both 30 clones plates and 2 clones (monoclonal) for the "a lot" plates, plus 1 "dirty take" on each plate. We used the Taq platinium protocol.
Cloning
We aimed to do the following cloning : - LacI-RBS2 -> E0240
- BB5 -> RO2
- Klenow fragment -> I13507
PCR
Digestion
Purification
Ligation
Transformation
Migration
People in the lab
Basile, Rafael, Nicolas
