Template:Team:Edinburgh/Notebook/Week05

ccm transformation
competent E coli were transformed with ccm plasmids

ccm transformants subculture
ccm transformed cells that lost RFP were subcultured.

liquid cultures preparation
subcultures of ccm transformed cells were used to inoculate 3ml LB; samples were left to grow overnight

primers preparation
Stock solution (500 pmol/ul, solution in EB) and working solution (10 pmol/ul 1 in 50 silution of stock solution in water) of primers were prepared for nfsl, cymA, mtrA and mtrCAB gene primers.

Kod PCR of Shewanella
Kod Pcr was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results.

Herculase PCR for Shewanella
Herculase PCR was performed for potential Shewanella oneidensis strain using 3 sets of primers: cymA, mtrA and mtrCAB. Gel analysis indicated no results.

Shewanella PCR Multiple PCRs were prepared:
Kod PCR for 16S ribosomal DNA for potential Shewanella strain (strains was later identified as different bacteria)
Taq PCR for 3 potential Shewanella strains using mtrA primers (one of the strains showed positive result at expected 1 kb)

Half-cell setup Half-cell was autoclaved, filled with liquid LB medium and inocluated with potential Shewanella strain. The Half-cell was sealed and left for incubation over the weekend. The initial voltage readout was 0.4 mV.