Labjournal

Week 2 (05/07 - 05/13/12)

weekly seminar:

Found our first sponsors: [http://corporate.evonik.com/en/Pages/default.aspx Evonik], [http://www.biocircle.com/en-ca/ BioCircle] and [http://www.merckgroup.com/en/index.html Merck], now treaties have to be created and signed
Julia is working on the database
Decision to organize waver sell to fill up our petty cash
Gabi and Isabel are designing a poster for the waver sell
For our human practices we wanted to find a sociology student, willing to think about bioethics, but failed
Our video is nearly done, is cutted and only needs be underlain with music

Team Modeling: Looking for suitable Software and enzymkinetics to model the degradation of our substrates with the different laccases. Finding the Michaelis-Menten kinetics and matlab.

Team Student Academy: First transformation of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 BBa_I13522] and plating on selective agar.
- Electroporation setup: U= 2,5 kV C= 25 µF and R= 400 Ω

Result: We got little colonies. There weren’t any green colonies and only some pale red fluorescent colonies.

Team Bacterial Laccases:

More PCRs of laccase genes CopA from Xanthomonas campestris B100 and CueO from E. coli BL21(DE3) with the isolated genomic DNA as template and Xcc_LAC_FW_T7 / Xcc_LAC_RV_HIS and B.pumi_LAC_FW_T7 / B. pumi_LAC_RV_HIS primer pairs.
Since we wanted to characterize laccases from different bacteria we had to order the bacterial strains which weren't available at the University Bielefeld from [http://www.dsmz.de/|DSMZ].

[http://www.dsmz.de/catalogues/details/culture/DSM-7039.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Thermus thermophilus HB27]

[http://www.dsmz.de/catalogues/details/culture/DSM-18197.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Bacillus halodurans C-125]

[http://www.dsmz.de/catalogues/details/culture/DSM-40069.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Streptomyces lavendulae REN-7]

[http://www.dsmz.de/catalogues/details/culture/DSM-40236.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Streptomyces griseus IFO 13350]

Team Student Academy: Repetition of the transformation didn’t change the result. We made a liquid culture of [http://partsregistry.org/Part:BBa_J04450 BBa_J04450], but it did not fluoresce. Searching for mistakes and alternatives. Maybe competent cells are not that good and in case of RFP the lacI sensitivity could be the problem.
Team Bacterial Laccases: After some empty agarose gels we finally isolated laccase gene CotA from Bacillus pumilus ATCC7061 as a PCR product. As template we used the plasmid we got from the Swiss working group.

Team Student Academy Testing the competent cells by transformation of pUC19. The transformation did not work that good, so that we produced new ones.

Team Bacterial Laccases PCR on Thermus thermophilus genomic DNA. First we dissolved some of the lyophilized powder in water and for opeing the cells we boiled them for a few minutes. The primers we used were T.thermo_LAC_FW_T7 and T.thermo_LAC_RV_HIS to get the laccase. Finally with aditional DMSO and GC-buffer we had a product of the GC-rich laccase.

=== Friday May 11th ===