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From 2012.igem.org

Contents 1 Labjournal 1.1 Week 4 (05/21 - 05/27/12) 1.1.1 Monday May 21st 1.1.2 Tuesday May 22nd 1.1.3 Wednesday May 23rd 1.1.4 Thursday May 24th 1.1.5 Friday May 25th 1.1.6 Saturday May 26th 1.1.7 Sunday May 27th

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 bts)

Monday May 21st

• Team Modeling: Our aims for modeling:

• • model the expression of the laccases in the organisms.
  • model the aktivity of our enzyms.
  • model the interesing parts of a clarification plant (the part witch are interesing for our cleaner.

• Team Bacterial Laccases: We did some restriction digests to clone our laccase PCR products from Xanthomonas campestris and E. coli in pSB1C3 backbone. Therefore we we transformed BBa_J04450 (pSB1C3 with RFP) in our competent KRX cells, picked colonies and made plasmidisolations. After that we ligated the cutted products with the cutted plasmid backbone.

Tuesday May 22nd

Team Bacterial Laccases: Transformation of the ligated plasmids in competent E. coli KRX cells.

Wednesday May 23rd

• Team Student Academy: Repetition of the transformation of BBa_J04450 and BBa_I13522 with new competent cells. For setup see here. Got intense red fluorescing colonies but no green fluorescing colonies. Made a backup of 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) plate we did the transformation with the same ligation preparation again. For the other ligation with pSB1C3 + Cueo (E. coli) we started colony PCRSs to find postive colonies. Sadly the colony PCR showed no product.
  • Purification of the T. thermo laccase PCR product out of agarose gel.

Thursday May 24th

• Team Student Academy: Made a liquid culture of BBa_J04450 at 30°C. There was no fluorescence. Furthermore BBa_J23100 was transformed. Also got intense red fluorescing colonies.

Friday May 25th

Team Bacterial Laccases:

• We had to to the PCR on T. thermo laccase again, because before the amount of purified PCR product was very low.

Saturday May 26th

Sunday May 27th