May

Milestones

• Initiating lab work on the sub-project endogenous mediator Glycerol dehydrogenase.
• Starting lab work with the first successful PCR: gldA with pre- and suffix overlaps could be amplified from Escherichia coli genome.
• The main work however is still planning of our sub-projects, designing experiments and a lot of research.
• Finding sponsors goes ahead. Many companies like our project and want to support us.

Week 1

Organization

• We were already able to find sponsors for our team: IIT BIEKUBA, IIT Biotech, Evonik, PlasmidFactory and FisherScientific will support us.

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Mediators

• Glycerol dehydrogenase
  • Primerdesign for isolation of gldA gene from Escherichia coli KRX strain, with overlaps for BioBrick prefix and suffix:
  • Forward primer gldA (43 bp): GAATTCGCGGCCGCTTCTAGATGGACCGCATTATTCAATCACC
  • Reverse primer gldA (45 bp): CTGCAGCGGCCGCTACTAGTATTATTCCCACTCTTGCAGGAAACG

2.Week

Organization

• Distribution of our team in various subgroups for best work efficiency.
• We’ve organized a barbecue to thank the working group of Dr. Jörn Kalinowski in the CeBiTec for the appropriation of space and support in the laboratory.

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3.Week

Organization

• Planning the trip to the European jamboree in Lyon from October 11th to 13th, 2013.

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Mediators

• Glycerol dehydrogenase
  • Starting first cultivation of Escherichia coli KRX strain for complete genome isolation.
  • Successful genome isolation of Escherichia coli KRX.
  • Successful PCR on the gldA gene of Escherichia coli KRX strain.

Table 1: Standard phusion PCR master mix.

Table 2: Two step standard phusion PCR program for gldA amplification.

• • The gldA PCR product was isolated by agarose gel electrophoresis and purified.
  • Bands are on at the expected size of 1050 bp.

Figure 1: Agarose gel from PCR on the gldA gene of Escherichia coli KRX strain with forward and reverse primer gldA. As a ladder we used [http://www.thermoscientificbio.com/nucleic-acid-electrophoresis/generuler-1-kb-dna-ladder-ready-to-use-250-to-10000-bp GeneRuler™ 1 kb DNA Ladder from Thermo Scientific].

4.Week

Organization

• We will also take part at the congress ‘‘Next generation of biotechnological processes 2020+‘‘ in Berlin on June 27th, 2013.
• Having a second presentation at Merck KGaA head office in Darmstadt for explaining our project in detail. We are happy that Merck will be our next supporter in 2013.

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