Team:Arizona State/Lab/Protocols/Extraction

Miniprep:

QIAprep kit using microcentrifuge:

• Place 1.5 mL liquid culture in Eppendorf tube.
• Spin down cells for 30sec-1min.
• Decant supernatant.
• Repeat steps 1-3.
• Resuspend pelleted cells in 250 uL Buffer P1.
• Transfer to microcentrifuge tube.
• Add 250 uL Buffer P2 – mix thoroughly by inverting tube 4-6 times.
• Add 350 uL Buffer N3 and mix immediately by inverting tube 4-6 times.
• Centrifuge for 10 minutes at 13,000 rpm.
• Add supernatant to QIAprep spin column.
• Centrifuge for 30-60sec. Discard flow-through.
• Add 500 uL Buffer PB, centrifuge 30-60sec. Discard flow-through.
• Add 750 uL Buffer PE, centrifuge for 30-60sec.
• Discard flow-through, centrifuge for additional 1 min, discard flow-through.
• Move spin column to new microcentrifuge tube. Add 50 uL Buffer EB to spin column.
• Let stand 1 min, centrifuge 1 min. Remove spin column. Label microcentrifuge tube.