Team:Berlin/Project
From 2014.igem.org
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</div> | </div> | ||
<div class="sub-content-project"> | <div class="sub-content-project"> | ||
- | <h2 class="sub-content-project-headline"> | + | <h2 class="sub-content-project-headline">Week 4: 21.04.2014 – 27. 04.2014 </h2> |
- | + | Minirep | |
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 5: 28.04.2014 – 04.05.2014</h2> | ||
+ | PCR Plasmid/Primar and Parameter check with Q5 Polymerase and Phusion Polymerase<br/> | ||
+ | <br/> | ||
+ | Miniprep of [pKD46+ DH5α] and [pKD4 + DH5 α]<br/> | ||
+ | Genomic DNA extraction from Pseudomonas putida<br/> | ||
+ | Enzyme digestion of different plasmids<br/> | ||
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 6: 05.05.2014 – 11.05.2014</h2> | ||
+ | Preculture of strains from Budisa strain databse in LB medium and Midiprep<br/> | ||
+ | <br/> | ||
+ | Canamycin cassette PCR of pKD4<br/> | ||
+ | <br/> | ||
+ | Restriction digest of Plasmid pKD4 and pKD46v<br/> | ||
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 7: 12.05.2014 – 18.05.2014</h2> | ||
+ | Biotransformation of Ferritin and pKD46 in Nissle and DH5 α strains | ||
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 8: 19.05.2014 – 25.05.2014</h2> | ||
+ | Gel extraction of FieF PCR<br/> | ||
+ | <br/> | ||
+ | Gene Knockout of FieF in [RV + pKD46] and [WM10+pKD46]<br/> | ||
+ | <br/> | ||
+ | Transformation of pKD46 in Nissle and MG 1655<br/> | ||
+ | <br/> | ||
+ | PCR of ATPCS and PPMT<br/> | ||
+ | <br/> | ||
+ | Digestion of pQE_80L for cloning<br/> | ||
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 9: 26.05.2014 – 01.06.2014</h2> | ||
+ | Production of LB plates<br/> | ||
+ | <br/> | ||
+ | AMB-1 and Microfluidic Chip were picked up from Max-Plank Institute<br/> | ||
+ | </div> | ||
+ | <div class="sub-content-project"> | ||
+ | <h2 class="sub-content-project-headline">Week 10: 02.06.2014 – 08.06.2014</h2> | ||
+ | Colony PCR and analytical gel electrophoresis for identifying the right clones<br/> | ||
+ | <br/> | ||
+ | PCR of ATPCS and PPMT to identify the correct amplification parameter.<br/> | ||
+ | <br/> | ||
+ | Colony PCR for Knockout.<br/> | ||
+ | <br/> | ||
+ | PCR of ATPCS and PPMT with Q5 High Fidelity Mastermix.<br/> | ||
</div> | </div> | ||
<div class="sub-content-project"> | <div class="sub-content-project"> |
Revision as of 21:34, 14 October 2014
Explore our Project:
What is it all about?
As previous iGEM teams have shown, synthesizing fully functional magnetosomes in E. coli is highly difficult as more than 60 highly regulated genes are involved. As a more feasible alternative, we simply want to synthesize magnetic nanoparticles in E. coli in order to attract cells with strong magnetic fields.
Therefore we want to use different strategies including manipulation of the iron homeostasis of E. coli, expression of different metal binding proteins such as ferritins and metallothioneins as well as a high-throughput growth medium optimization.
Furthermore, we will work with other metal binding proteins such as metallothioneins and phytochelatin synthases in order to achieve nanoparticle synthesis.
Once we have discovered the best way to magnetize E. coli bacteria, we will build and characterize suitable BioBricks that can be used by any research lab or iGEM team in the world in order to remote control the cellular movement.
Lab Summary
Week 1: 02.04.2014 – 06.04.2014
Cultivation of E. coli Nissle 1917Genomic DNA extraction of E.coli Nissle strain
Production of BfR, FTNA1 and FTNA2
Restriction digest
PCR Purification
Ligation Assay
Transformation into DH5α-Cells
Week 2: 07.04.2014 – 13.04.2014
Colony PCR to check the resultsBfr, FTNA 1, FTNA 2 primar designed for amplification
Transformation of pQE_80L into DH5α
Cultivation of BfR, FTNA 1, FTNA 2 in LB
Week 3: 14.04.2014 – 20.04.2014
Miniprep of the cells from Week 2DNA concentration determination and sequencing
Expression of BfR, FTNA 1, FTNA 2 and induction with IPTG
Production of a Mutaflor-Supression culture and streaking
Week 4: 21.04.2014 – 27. 04.2014
MinirepWeek 5: 28.04.2014 – 04.05.2014
PCR Plasmid/Primar and Parameter check with Q5 Polymerase and Phusion PolymeraseMiniprep of [pKD46+ DH5α] and [pKD4 + DH5 α]
Genomic DNA extraction from Pseudomonas putida
Enzyme digestion of different plasmids
Week 6: 05.05.2014 – 11.05.2014
Preculture of strains from Budisa strain databse in LB medium and MidiprepCanamycin cassette PCR of pKD4
Restriction digest of Plasmid pKD4 and pKD46v
Week 7: 12.05.2014 – 18.05.2014
Biotransformation of Ferritin and pKD46 in Nissle and DH5 α strainsWeek 8: 19.05.2014 – 25.05.2014
Gel extraction of FieF PCRGene Knockout of FieF in [RV + pKD46] and [WM10+pKD46]
Transformation of pKD46 in Nissle and MG 1655
PCR of ATPCS and PPMT
Digestion of pQE_80L for cloning
Week 9: 26.05.2014 – 01.06.2014
Production of LB platesAMB-1 and Microfluidic Chip were picked up from Max-Plank Institute
Week 10: 02.06.2014 – 08.06.2014
Colony PCR and analytical gel electrophoresis for identifying the right clonesPCR of ATPCS and PPMT to identify the correct amplification parameter.
Colony PCR for Knockout.
PCR of ATPCS and PPMT with Q5 High Fidelity Mastermix.