Team:Bielefeld-Germany/Labjournal/June

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June

Milesstones





5.Week

Organization

  • iGEM-Team Bielefeld will support the ‘CeBiTec Pupil Academy’ from 26.-30. August with an own experiment.

MFC

Mediators

Cytochromes

Biosafety

Porines









6.Week

Organization

  • Thanks to NEB Biolabs for the free iGEM kit with many useful laboratory things for all German iGEM teams.
  • We are working on our first press release.
  • Having a short radio contribution in the Bielefeld university campus radio (radio 87.9 hertz).


MFC

Mediators

Glycerol dehydrogenase
  • Isolation of shipping vector pSB1C3 out of 2013 Distribution Kit Plate 5 Well 3A with insert Part RFP (J04450) for better transformation characterization (Distribution Kit BioBrick isolation).
  • Transformation of BBa_J04450 into Escherichia coli KRX strain.
  • Plasmid isolation of BBa_J04450.


Cytochromes

Biosafety

Porines












7.Week

Organization

  • Nothing organizational.

MFC

Mediators

Cytochromes

  • Cultivation of Shewanella oneidensis MR-1 in liquid LB medium at 30 °C


  • Isolation of genomic DNA from S. oneidensis
    • 4-2006-451: Conc.:453.1 ng/µl OD260/280: 1.92 OD260/230:2.78
    • 4-2006-452: Conc.:447.1 ng/µl OD260/280: 1.92 OD260/230:2.67
    • Dilution to PCR template: 4-2006-453: 5.5ng/µl


  • Amplification of the mtrCAB cluster
    • Gradient: 55.8-56.7-57.8-59.1-60.4-61.7-62.9-63.9
    • Clear Bands at the expected 5.2kb


  • PCR-CleanUp
    • PCR-cleanup of No.2 and No.5 with Macherey-Nagel-Kit and NanoDrop-Measurement
    • Lane2: 4-2106-451: 7.4 ng/µl
    • Lane5: 4-2106-451: 8.5 ng/µl


Biosafety

Porines











8.Week

Organization

  • Let’s go to Lyon, flights are booked for the European jamboree in Lyon from 11.-13. October 2013.
  • We will participate at ‘BioNRW pHD Student Convention’ in Düsseldorf at 13. July.

MFC

Mediators

Glycerol dehydrogenase
Figure 1: Agarosegel with NEB 1 kb DNA Ladder as marker. Bands are showing restriction analysis from cloning of GldA into pSB1C3 shipping vector with NEB Biobrick assembly Kit. Assembly did not work, only one band at the size of 2000 bp showing religated pSB1C3.
  • Cloning of GldA into pSB1C3 shipping vector with NEB Biobrick assembly Kit did not work as expected.
  • Screening of colonies with colony PCR and Plasmid restriction analysis shows religated pSB1C3 shipping vector.
  • Bands are at size of 2000 bp, the length of linear pSB1C3.
  • Primerdesign for pSB1C3 according an universal usable backbone for Gibson Assembly with Prefix and Suffix specific overlaps:
  • Forward Primer pSB1C3 (23 bp): TACTAGTAGCGGCCGCTGCAGTC
  • Reverse Primer pSB1C3 (23 bp): CTCTAGAAGCGGCCGCGAATTCC


Cytochromes

Biosafety

Porines












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