"
Team:SDU-Denmark/labwork/Notebook/week8
From 2012.igem.org
| Line 299: | Line 299: | ||
<p> | <p> | ||
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 | 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. | + | and incubated overnight at 37C.<br/><br/> |
| - | The following day we had colonies for both. These were transfered to liquid medium. | + | The following day we had colonies for both. These were transfered to liquid medium.<br/><br/> |
| - | 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 | + | 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.<br/><br/> |
| - | 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. | + | 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.<br/> |
</p> | </p> | ||
Revision as of 02:14, 27 September 2012
Laboratory Notebook
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.
