Team:Bielefeld-Germany/Labjournal/week4

Revision as of 13:25, 13 September 2012 by Juvoss (Talk | contribs)

Week 4 (05/21 - 05/27/12)

weekly seminar:

Lab service: Isabel
We try to establish a collaboration with the iGEM team from SDU-Denmark
Got our distribution kits
First successful cloning and cultivations
Who wants to be a summer school teacher?
We will not travel to the ACHEMA because only local teams are invited
Do we want to participate in the Biolympics? (It's a sports party with fun organized by the [http://bts-ev.de/ bts])

- model the expression of the laccases in the organisms.
- model the activity of our enzymes.
- model the interesting parts of a clarification plant (the part witch are interesting for our cleaner.

Team Bacterial Laccases: We wanted to clone our laccase PCR products from Xanthomonas campestris and E. coli in pSB1C3 backbone. Therefore we did some restriction digests on the PCR products and the vector [http://partsregistry.org/Part:BBa_J04450 BBa_J04450].

Team Bacterial Laccases: Ligation of the digested PCR products in pSB1C3 backbone and transformation in KRX electro-competent cells.

Team Student Academy: Repetition of the transformation of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 BBa_I13522] with new competent cells. For setup see here. Got intense red fluorescing colonies but no green fluorescing colonies. Made a backup of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450]. Asking for other plasmids containing GFP at the working groups of our University.

Team Bacterial Laccases:
- After there were no colonies on our pSB1C3 + CopA (Xanthomonas campestris) transformation plate we did the transformation with the same ligation preparation again. The other ligation with pSB1C3 + CueO (E. coli) showed colonies so we started colony PCRs to find positive colonies. Sadly the colony PCRs showed no products but the problem was that we just had the long overhang primers (E.coli_LAC_FW_T7 and E.coli_LAC_RV_HIS ). Therefore we ordered the F and R primers.

Team Student Academy: Made a liquid culture of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450] at 30°C. There was no fluorescence. Furthermore [http://partsregistry.org/Part:BBa_J23100 BBa_J23100] was transformed. Also got intense red fluorescing colonies.

Team Bacterial Laccases:

We had to do the PCR on T. thermo laccase again because after the purification of the last PCR product the DNA amount was very low.

Sunday