EPF-Lausanne/26 August 2009

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Revision as of 09:12, 4 September 2009

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

PCR

Digestion

Purification

Ligation

Transformation

Migration

People in the lab

Basile, Rafael, Nicolas

@@ Line 34: / Line 34: @@
 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.
+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===