Team:Bielefeld-Germany/Labjournal/week4
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Labjournal
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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])
Monday May 21st
- Team Modeling: Our aims for modeling:
- 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].
Tuesday May 22nd
Team Bacterial Laccases: Ligation of the digested PCR products in pSB1C3 backbone and transformation in KRX electro-competent cells.
Wednesday May 23rd
- 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.
- Purification of the T. thermo laccase PCR product out of agarose gel.
Thursday May 24th
- 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.
Friday May 25th
Team Bacterial Laccases:
- We had to do the PCR on T. thermo laccase again, because before the amount of purified PCR product was very low.