Team:Lyon-INSA-ENS/Realisation/Protocols/DNA-purification

Plasmid or Cosmid DNA Purification Using HiSpeed Plasmid Midi Kits

This protocol is for preparation of up to 200µg of high- or low-copy plasmid or cosmid DNA using the HiSpeed Plasmid Midi Kit.

A final DNA concentration of up to 0.4µg/µL can be expected, if eluting a high-copy plasmid with 500µL of Buffer TE.

Procedure

1. Pick a single colony from a freshly streaked selective plate and inoculate a starter culture of 2-5mL LB meduim containing the appropriate selective antibiotic. Incubate for approx. 8h at 37°C with vigorious shaking (approx. 300 rpm).

2. Dilute the starter 1/500 to 1/1000 into selective LB medium. For high-copy plasmids inoculate 50mL. Grow at 37°C for 12-16h with vigorous shaking (approx. 300 rpm).