Team:SDU-Denmark/labwork/Notebook/week8

From 2012.igem.org

iGEM TEAM ::: SDU-DENMARK

Laboratory Notebook

1st week 2nd week 3th week 4th week
5th week 6th week 7th week 8th week
9th week 10th week 11th week 12th week

20-08-2012 to 26-08-2012

We recieved the Shine-dalgarno inducing primers and mutagenesis were performed on 1-SST and 1-FFT cultures, XL10-GOLD bacteria were transformed with these and incubated overnight at 37C.

The following day we had colonies for both. These were transfered to liquid medium.

The minipreps from the liquid cultures the following days yielded unexpectedly low results in the area around and below 10ng/uL. We were unsure why this occoured, but we began to expect the genes, which now contained bacterial RBS might be greatly inhibiting the bacterias growth. This does make sense, since inulin synthhesis would compete for sucrose with the bacterias sucrose metabolism, plus the synthesized inulin might be building up inside the bacteria. If this was the case, we needed to find a way around it in order to continue working efficiently with our genes.

We did not have anything available that would be able to inhibit the pJET promoter, so we took a more simple approach: increase the incubation time with more medium.