Team:RHIT/Protocol

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Protocols

Pellet 1-5 ml bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature.
Resuspend pelleted bacterial cells in 250 µl Buffer P1 and transfer to a microcentrifuge tube.
Add 250 µl Buffer P2 and mix thoroughly by inverting the tube 4-6 times until the solution becomes clear. Do not allow the lysis reaction to proceed for more than 5 min.
Add 350 µl Buffer N3 and mix immediately and thoroughly by inverting the tube 4-6 times.
Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge.
Apply the supernatant from step 5 to the QIAprep spin column by decanting or pipetting. Centrifuge for 30-60 s and discard the flow-through.
Recommended: Wash the QIAprep spin column by adding 500 µl Buffer PB. Centrifuge for 30-60 x and discard the flow-through.
Wash the QIAprep spin column by adding 750 µl Buffer PE. Centrifuge for 30-60 s and discard the flow-through. Transfer the QIAprep spin column to the collection tube.
Centrifuge for 1 min to remove residual wash buffer.
Place the QIAprep column in a clean 1.5 ml microcentrifuge tube. To elute DNA, add 50 µl Buffer EB or water to the center of the QIAprep spin column, let it stand for 1 min, and centrifuge for 1 min.