Griffitts:PCR clean-up
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Materials
- Buffer PB (in Qiagen kit)
- Buffer PE (in Qiagen kit)
- Buffer EB (in Qiagen kit)
- QiaPrep spin column (1 per PCR reaction)
- 1.5 mL microcentrifuge tube (1 per PCR reaction)
Procedure
- Label columns and microcentrifuge tubes with the names of the appropriate PCR reactions
- Using P-200, add 200 μL buffer PB to each 40-μL PCR and mix well by gently pipeting up and down
- Transfer mixture to a QiaPrep spin column
- Centrifuge at full speed for 30 seconds
- Dump and tap
- Add 600 μL buffer PE to each column
- Centrifuge at full speed for 30 seconds
- Dump and tap
- Centrifuge for an additional 30 seconds at full speed
- Move column to a 1.5 mL microfuge tube
- Add 50 μL TE buffer (to elute)
- Drip this directly onto the filter disk but don't touch it with the pipette tip
- Wait 1 minute
- Centrifuge at full speed for 1 minute
- Point the lids of the microcentrifuge tubes in a clockwise direction, otherwise they'll break off