Labjournal

Week1 Week2 Week3 Week4 Week5 Week6 Week7 Week8 Week9 Week10 Week11 Week12 Week13 Week14 Week15 Week16 Week17 Week18 Week19 Week20 Week21

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 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.

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 after the purification of the last PCR product the DNA amount was very low.

Saturday May 26th

Sunday May 27th