For the isolation following kit has been used: Fermentas GeneJET™ Plasmid Miniprep Kit

- Centrifuge 10 mL of over-night liquid culture

- Resuspend pellet in 800 µL of resuspension solution

- Ribolyse three times for 60 s with 6500 rpm (30 s break between every run, 400 mg beads)

- Centrifuge 3 min at 10,000 rpm

- Transfer 500 µL of supernatant into new 2 mL tube

- Add 500 µL of lysis buffer, invert 4 - 6 times

- Add 700 µL of neutralisation buffer, invert 4 - 6 times

- Centrifuge 10 min at 12,000 rpm

- Transfer 2 x 600 µL of supernatant into spin column (centrifuge down each time for 30 s at 12,000 rpm)

- Two wash steps with 750 µL of washing buffer

- Dry column

- Elute in 75 µL of elution buffer