Team:Aberdeen Scotland/Mini Preps

Mini Preps
Our mini preps were prepared using QIAprep Spin Miniprep Kits from Qiagen. The protocol is from the handbook, using a Microcentrifuge.

Protocol
1. Culture E.coli overnight in 5ml LB medium in a shaking incubator at 37C 2. Centrifuge culture at 4000 rpm for 10 minutes. 3. Resuspend pelleted bacterial cells in 250µl Buffer P1 and transfer to a microcentrifuge tube. 4. Add 250µl Buffer P2 and mix thoroughly by inverting the tube 4-6 times. 5. Add 350µl Buffer N3 and mix immediately and thoroughly by inverting tube 4-6 times. 6. Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge. 7. Apply supernatant to QIAprep spin column by decanting/pipetting. 8. Centrifuge for 60s at 13,000 rpm and discard flow-through. 9. Wash column by adding 0.5ml Buffer PB, centrifuge for 60s and discard flow-through. 10. Wash column by adding 0.75ml Buffer PE, centrifuge for 60s and discard flow-through. 11. Centrifuge for an additional minute and discard flow-through. 12. To elute DNA, place QIAprep column in clean 1.5ml microcentrifuge tube. Add 50µl Buffer EB (or water) to center of each QIAprep spin column, let stand for 2 min and centrifuge for 1 min.