Latest revision as of 08:53, 10 September 2009

Project progress

Restriction digest with EcoRI and PstI on and the purified pcr product of in order to get our promotor on itself in the standard vector.
Gel electroforesis and extraction of this RD. The concentration of this extraction was too low so no ligation was performed with these samples.
Extra pcr was done on with primers 2260 and 2261.
Liquid cultures of LigX and of were made from the 4x plates.

Miniprep of the four colonies from EF ligation
- Accidentally used double volumes of elution buffer on EF4

part	concentration	260/280	260/230
EF 1	115,5	1,98	2,31
EF 2	105,2	1,96	1,66
EF 3	95,1	2,05	0,98
EF 4	52,7	2,06	0,97

Gel electrophoresis from restriction on sam5 and sam8 looked good
Purified and ligated the restriction from sam5 and sam8 overnight
- Concentration of sam8 was barely enough for ligation...

part	concentration	260/280	260/230
sam5	10,1	2,11	0,04
sam8	3,1	3,96	0,02

a new restrictiondigest was done for puc and A because we decided that earlier results were not good enough.
There was a gelextraction, nanodrop and ligation done overnight.
The colony PCR of rpoA => 428 again for one minute => bad result
Miniprep of X+K was checked and put on gel. Wrong result probably because we selected a wrong colony.

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