Team:KULeuven/27 August 2008

Wet Lab
We continued our ligations. Therefore we miniprepped and digested the following parts: C0040+B0015, C0012+B0015, K145015+B0015, K145001+B0015, K145151+B0015 (with XbaI) and R0040+J23022 (with EcoRI and SpeI). However, when we put the digests on an agarose gel, we didn't see anything. But, according to the nanodrop, there should be DNA in our samples. So we put the miniprepped plasmids on gel (not digested) and we couldn't see anything - again. This means that either the miniprep failed or the colonies were crap. Tomorrow we will electroporate the ligated parts again (but we will plate them on Ap instead of Km). We will also redo the ligations, but this time we will use the protein coding regions as vector and B0015 as insert. This means cutting the vectors with SpeI and PstI and B0015 with XbaI and PstI.

Modelling has shown that our original setup requires some (major) adaptations. We miniprepped and digested some of the parts needed for these changes: B0033, B0034 and C0012. We also did this for R0011 and F1610, but F1610 seemed to be degraded - again.

There were colonies on the plates with K145001+pSB1A2 and K145151+pSB1A2. Liquid cultures were made.

We will make a glycerol stock of the ligations that succeeded. Therefore, we made a liquid culture of J23116+B0032, R0040+B0032, I712074+J23032, J23109+J23032and R0040+J23022.

Ethics
Worked some more on the ethics-part. Did some more research.