QIAquick PCR Purification Kit Microfuge Protocol

Ensure 100% ethanol has been added to PE buffer before starting extraction. Volume to add is on the bottle label.

Manipulations should be done with post-PCR pipettes and tips

  1. Add 5 volumes of PB buffer to 1 volume of PCR sample and vortex. If the samples are in 0.2ml tubes, transfer them to 0.6 or 1.5ml tubes and rinse the original 0.2ml tubes out with the PB buffer.
  2. Place a labelled QIAquick column in a labelled collection tube.
  3. Add each sample to the corresponding column, spin for 1 minute.
  4. Discard flow through into 5% Virkon, replace column into collection tube.
  5. Add 0.75ml PE buffer to each column and spin for 1 minute.
  6. Discard flow through into 5% Virkon, respin for 1 minute. Discard collection tubes.
  7. Place each column in a labelled 1.5ml tube.
  8. Add 30ml of BDH water to the centre of the column and leave to stand for 1 minute, then spin for 1 minute.
  9. Measure DNA concentration, store samples at -20C or lower.