Team:Evry/Notebook/w11

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Iron coli project

Week 11: 26th August - 1st September

TECAN

Creation of a E. Coli ΔFur

Strain preparation

Chemically competent Top 10 E. Coli is transformed with the plasmid PTK which contain λ Red factor. We plated our bacteria on LB Agar with spectinomycin and let them grow overnight at 30°C. One colony had grown on our plate. We started an overnight culture of this colony in 2 mL of LB with spectinomycin and still at 30°C. We then diluted 1 mL of our culture in 100 mL of LB with spectinomycin and IPTG so the λ Red factor would start to be expressed. We let bacteria grew to a OD of 0.5 and made them electro competent.

Electro competent