Mini prep- Plasmid DNA isolation (manual method)

•Overnight grown culture harvested by centrifugation at 8000 for 2min

•Pellet was resuspended in 250µl of P1 Buffer by vortexing

•250µl of Buffer P2 was added followed by gentle invert mixing (4-6 times) and kept for 2-3 min exact

•350µl of Buffer P3 was added and mixed gently by inverting

•Supernatant was collected in a fresh microfuge tube and 520µl of iopropanol was added to it

•Incubated for 20 min followed by centrifugation at 13000 rpm for 15 min

•Supernatant was discared and 800µl of chilled 70% ethanol was added and mixed well

•Centrifuged at 13000rpm for 2 min

•The pellet was air dried and resuspended in 30µl of AMQ at 55ºC

•Stored at -20ºC