Team:Northwestern/pure
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Revision as of 23:59, 27 September 2013
PCR Purification
We used the QIAquick PCR Purification Kit from Qiagen.
- Add 5 volumes of Buffer PB to 1 volume of PCR sample and mix. Apply sample to a spin column and centrifuge at 13,000 rpm for 30-60 seconds.
- Discard flow-through.
- Place the column back into the same tube.
- Wash with 0.75 mL Buffer PE and centrifuge at 13,000 rpm for 30-60 seconds.
- Discard flow-through and place the column back into the same tube.
- Dry spin for an additional 1 minute at 13,000 rpm.
- Place column in a clean 1.5 mL microcentrifuge tube. Add 50 uL of nuclease free water to the center of the membrane.
- Let sit at room temperature for 10 minutes.
- Centrifuge the column at 13,000 rpm for one minute to elute.
- Store at -20°C.