Team:Bielefeld-Germany/Labjournal/week2

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(Monday May 7th)
(Week 2 (05/07 - 05/13/12))
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==Week 2 (05/07 - 05/13/12)==
==Week 2 (05/07 - 05/13/12)==
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* '''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.
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* '''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.
=== Monday May 7th ===
=== Monday May 7th ===

Revision as of 08:18, 5 September 2012

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 substrates 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 and 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.

Friday May 11th