Team:KULeuven/7 August 2008

Wet Lab


We prepared a digest of F1610 and R0062. F1610 wasn't cut with XbaI. We are doing this again. The result will follow tomorrow.

The primers finally arrived and we did a PCR on the ligated parts with primers VR and VF-2. This way we could see whether the ligations succeeded or not.

We also set up some ligations today: C0012+B0015, R0062+B0032, R0084+J23022.

The cells we transformed yesterday all gave colonies. We streaked them out and made a fluid culture. The digests we made yesterday were purified from gel.

General
Cell death contains an error: P1010 is a part which constitutively produces ccdB. It already contains a promoter and a RBS, see the parts page and the Hard Information. Primers will have to be made to create a full-blown biobrick with just the ccd locus. A new part (K145151) has been made from P1010 with just the ccdB coding sequence. We will PCR this part starting from P1010 with the correct primers.

Polishing the hybrid promoter and checking the promoters in the memory.

Modeling
We continued our work on the sensitivity analyses. Things are looking quite ok. We could see that the output of the filter could be. Only in the last few minutes we got some weird results. Will look into it tomorrow.

The memory was split wide open again. We're now using a non-LVA tagged 434 cI downstream of the OmpF promoter in order to shorten the length of the lightpulse required to switch to the ON state. The lifetime of this cI is estimated to be about 2 hours. This will require a lot of tinkering on the memory again.