Labjournal

Week 1 (04/30 - 05/06/12)

Start of our WET LAB time.
Generating new competent E.coli KRX cells.
Cultiviation of Xanthomonas campestris B100 and E. coli BL21(DE3). The bacterial strains we got from a working group at our University. After cultivation we isolated the genomic DNA. The DNA was needed as template for PCRs to purify the wanted laccase ORFs.
Sending requests to working groups for different plasmids , which have already worked with laccases and described them in their papers. Unfortunately just one answer came back (thanks a lot to them).So we got a vector with the laccase-ORF CotA from Bacillus pumilus ATCC7061 from the Swiss Federal Laboratories for Materials Science and Technology, Laboratory for Biomaterials in Switzerland.

Team Student Academy: We got the chance to organize one part of the first school academy “synthetic biology/ biotechnology” at the CeBiTec of University Bielefeld by arranging experiments for the pupils and by presenting us and the iGEM competition. For the experimental part our general idea was to give them an understanding of principle methods in biotechnology / synthetic biology by using fluorescent proteins. The planned experiments:
- Plasmidisolation of RFP/GFP from a liquid culture.
- Transformation of a plasmid mixture consisting of two different fluorescent proteins (e.g. RFP and GFP) and different antibiotic resistances into E.coli KRX. It will be plated out on LB agar plates without antibiotics and on plates containing one of the two antibiotics, which are present on the plasmids. This way we can demonstrate the effect of antibiotics as selective pressure.

Team Bacterial Laccase: After the vector has arrived, we transformed it into the competent E.coli KRX which we have already generated. The protocol we used was as followed:
- The electroporation setup: U= 2,5kV C= 25 µF and R= 400 <math>\omega</math>
- Since we did not know the efficient of our competent KRX we used two different E.coli volumes for the transformation, 50µL and 100µL. We gave 50µL 10% Gylcerol to the reaction tubes with 1µL of the vector DNA (Bacillus pumilus). After the transformation we plated them into ampicillin plates.
Team Bacterial Laccase: PCR with the Xanthomonas campestris B100 and E. coli BL21(DE3) genomic DNA.
- PCR table

Cycle between step 2 and 4 35 times.

weekly seminar:

Do we want to order strains of Trametes versicolor and Trametes villosa?
Gathering information about signal sequences in yeast
Decision to create a database, so that we can easily number and inscribe our lab results
Decision to arrange a summer school for pupils in their last year before the final exams
Discussion about how to meet a member of the german Bundestag (the german parliament)