Team:Evry/Notebook/Transposons/15.10.14

== 14.10.2014 ligation== it gave only one clone. Clone culture: 5 ml LB + 5 ul Kan (25 mg/ml) - 37 C° == Transformation Pseudovibrio denitrificans & Dh5 alpha with mRFP in BBa_1413044== electrocompetent cells: 1 mm cuvette; 1800 V recuperation media: MB for Pseudovibrio denitrificans , LB for DH5alpha plating: MB agar for Pseudovibrio denitrificans LB agar for LB for DH5alpha == Transformation Pseudovibrio denitrificans & Dh5 alpha with BBa_1413001 in BBa_1413044== electrocompetent cells: 1 mm cuvette; 1800 V recuperation media: MB for Pseudovibrio denitrificans , LB for DH5alpha plating: MB agar for Pseudovibrio denitrificans LB agar for LB for DH5alpha == Verification testing of pnK2 in Pseudovibrio denitrificans 16 clones of Pseudovibrio denitrificans were grown in 3 ml MB, 30 C°, 24h DNA extraction: Invitrogen DNA extraction kit used with the Gram - protocol. nanodrops gave value between 80 ng/ul & 130 ng/ul PCR for Kan gene insertion: PCR: enzyme: Q5; template: 200 ng genomic preparation Pseudovibrio denitrificans; oligo: F 82/ R 83; Tm tested: 55; elongation time: 2m00s PCR for Pseudovibrio denitrificans specific oligo: PCR: enzyme: Q5; template: 200 ng genomic preparation Pseudovibrio denitrificans; oligo: F 72/ R 73; Tm tested: 55; elongation time: 2m00s