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Team:Bielefeld-Germany/Labjournal/July
From 2013.igem.org
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Revision as of 21:57, 20 September 2013
Milestones
1.Week
Organization
- We’ve presented us at the congress ‘Next generation of biotechnological processes 2020+’ in Berlin at 27. June 2013. There we have found an expert (Dr. Falk Harnisch) on the topic of MFC and we have decided to participate the German Synthetic Biology Day.
MFC
Mediators
Cytochromes
The mtrCAB fragment has two illegal PstI restriction sites at x xbp and yy bp, so we had to design new primers to remove them. We replaced one base in each restriction site, without affecting the coding triplett, by respective primer overlaps . We ended up with three different fragments, which will be ligated back together via Gibson Assembly.
mtrCAB_Frag1_rev:
mtrCAB_Frag2_fwd:
mtrCAB_Frag2_rev:
mtrCAB_Frag3_fwd:
Biosafety
Porines
2.Week
Organization
- We can present new sponsors of iGEM-Team Bielefeld: Stockmeier, Baxter, Promega, BIO.NRW and Applichem will support our team.
MFC
Mediators
Cytochromes
Biosafety
Porines
3.Week
Organization
- We’ve presented us at the congress “BioNRW pHD Student Convention” in Düsseldorf at 13. July 2013 with a short presentation about iGEM and our project.
- We are proudly to present our first press release it was amazing to see how often our press release was picked of.
MFC
Mediators
Cytochromes
Biosafety
Porines
4.Week
Organization
- Dr. Falk Harnisch will have a presentation at CeBiTec colloquium as an expert for our team.
- Preparing experiments for the ‘Day of Synthetic Biology’. Our ideas are to have different experiments like DNA isolation from fruit and vegetables, pipetting of bright colors, chromatography with markers, a potato battery or microscopy.
- The TV station WDR would like to contribute with us on our topic
MFC
Mediators
Cytochromes
Size: 1800 bp
Program: PhusionPCR
Gradient: 54°C - 71°C over 8 steps
Primer: mtrC_fwd & mtr_Frag1_rev
Template: S. oneidensis PCR Template from genomic DNA
Notes: Annealing temperature has no significant impact on the PCR
Size: 330 bp
Program: PhusionPCR
Primer: mtr_Frag2_fwd & mtr_Frag2_rev
Template: S. oneidensis PCR Template from genomic DNA
Notes:
Size: 3000 bp
Program: PhusionPCR <
Gradient: 54°C - 71°C over 8 steps
Primer: mtr_Frag3_fwd & mtrB_rev
Template: S. oneidensis PCR Template from genomic DNA
Notes: Annealing temperature has no significant impact on the PCR
Size:6311 bp
Program: PhusionPCR <
Gradient: 51.5°C - 68°C over 8 steps
Primer: ccmAH_fwd & ccmAH_rev
Template: E. coli PCR Template from genomic DNA
Notes: A high annealing temperature of 68°C or more increases yield as well as reduces unspecific bands and is therefore recommended.
Gelextraction and clean up with the AnalytikJena GelExtraction-Kit-Elution buffer was preheated to 50°C
-We eluted each column twice, with 20 ul each and combined it afterwards
Measurement of nucleid acid concentration via NanoDrop-Fragment1: 4-2607-304: 10.5 ng/ul
-Fragment2: 4-2707-003: 44.6 ng/ul
-Fragment2: 4-2707-004: 13.5 ng/ul
-Fragment3: 4-2607-301: 11.1 ng/ul
-Fragment3: 4-2607-302: 8.9 ng/ul
-Fragment3: 4-2607-303: 9.2 ng/ul
-ccmAH: 4-2807-302: 11.0 ng/ul
-ccmAH: 4-2807-303: 11.5 ng/ul
Biosafety
Porines
5.Week
Organization
- Spontaneous visit of Radio Bielefeld in our laboratory. In addition to a radio interview a short video clip was filmed
