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Team:Bielefeld-Germany/Labjournal/week2
From 2012.igem.org
Contents |
Labjournal
Week1 Week2 Week3 Week4 Week5 Week6 Week7 Week8 Week9 Week10 Week11 Week12 Week13 Week14 Week15 Week16 Week17 Week18 Week19
Week 2 (05/07 - 05/13/12)
- Team Modeling: Looking for suitable Software and enzymkinetics to model the degradation of our substartes with the different laccases. Finding the Michaelis-Menten kinetics and matlab.
Monday May 7th
- 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
| Material | Volume |
|---|---|
| E. coli KRX competent cells | 50 µL |
| glycerol (10 %) | 50 µL |
| plasmid | 1 µL |
- Result: We got little colonies. There weren’t any green colonies and only some pale red fluorescent colonies.
Team Bacterial Laccases:
- Successful PCRs of laccase genes CopA from Xanthomonas campestris B100, CueO from E. coli BL21(DE3) with the isolated genomic DNA as template.
- Because we want to characterise 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]
Tuesday May 8th
- 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.
Wednesday May 9th
Thursday May 10th
- Team Student Academy Testing the competent cells by transformation of pUC19. The transformation did not work that good, so that we produced new ones.
