Alkaline Lysis Miniprep Using QIAGEN QIAprep Kit

1. Alkaline Lysis Miniprep Using QIAGEN QIAprep Kit Remove 1ml of overnight culture and place in a clean, sterile 1.5 ml eppendorf tube. Centrifuge for 5 minutes at 14,000 rpm in a microcentrifuge. Discard supernatant. Add 250 ml of Buffer P1 to the pelleted cells. Resuspend. Add 250 ml of Buffer P2 to the pelleted cells. Mix thoroughly but gently. Add 350 ml of Buffer N3 to the pelleted cells. Mix thoroughly but gently. Centrifuge for 10 minutes at 14,000 rpm in a microcentrifuge. Add supernatant to QIAprep spin column. Centrifuge for 1 minute at 14,000 rpm. Discard flow-through. Wash QIAprep spin column by adding 750 ml of Buffer PE. Centrifuge for 1 minute at 14,000 rpm. Discard flow-through. Centrifuge for 1 minute at 14,000 rpm. Discard flow-through. This removes residual buffer. Place the QIAprep spin column into a clean, sterile 1.5 ml eppendorf tube. Add 50 ml of sterile dH2O to the center of the QIAprep spin column. Let stand for 1 minute. Centrifuge for 1 minute at 14,000 rpm. Discard the QIAprep spin column. Retain the DNA miniprep for further analysis. Store at -20oC.