Team:Bielefeld-Germany/Labjournal/June
From 2013.igem.org
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
- Primerdesign for isolation of OprF from Pseudomonas fluorescens strain, with overlaps for Biobrick Prefix and Suffix:
- Forward Primer OprF (49 bp): GAATTCGCGGCCGCTTCTAGATGAAACTGAAAAACACCTTGGGCTTTGC
- Reverse Primer OprF (51 bp): CTGCAGCGGCCGCTACTAGTATTACTTAGCTTGGGCTTCAACCTGCGCTTC
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
File:Bielefeld2011 BPAAbbau Versuch 1.jpg
Figure 1: Agarosegel from PCR on the OprF gene of Pseudomonas fluorescens strain with forward and reverse primer OprF. For Ladder we used GeneRuler™ 1 kb DNA Ladder fromThermo Scientific.
- Starting first cultivation of Pseudomonas fluorescens strain for complete genome isolation.
- Successful genome isolation of Pseudomonas fluorescens.
- Successful PCR with Forward and Reverse Primer OprF on the OprF gene of Pseudomonas fluorescens strain.
- OprF PCR product was isolated by Agarose gel electrophorese and purificated.
- Bands are at expected size of 1300 bp.
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
- 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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