Modified from Qiagen’s protocol with optimizations for low concentration PCR products

Works for products >100bp. Products as small as 50bp may work but concentrations will be reduced.

  1. Run 2uL of PCR product on a minigel to ensure the reaction worked.
  2. In a 1.5uL tube mix 5x the PCR product volumes of Buffer PB with 1uL pH indicator per 250uL Buffer. Mix and observe yellow color.
  3. Add the PCR product and mix by pipetting.
  4. If color changes from yellow to orange or purple, add 10uL 3M sodium acetate and mix.
  5. Place in a QIAquick column and provided 2mL collection tube.
  6. Centrifuge (max speed) 1 minute.
  7. Discard flow-through and place the column back in the collection tube.
  8. To wash, add 750uL Buffer PE to the column.
  9. Centrifuge 1 minute.
  10. Discard flow-through and place the column back in the collection tube.
  11. Centrifuge 1 minute to remove residual wash buffer.
  12. Discard collection tube and place column in labeled 1.5mL tube.
  13. To elute DNA, add 35uL Buffer EB to the center of the column membrane.
  14. Let column stand 2 minutes to maximize yield.
  15. Centrifuge 1 minute.
  16. Discard column. Purified PCR product is in the 1.5mL tube.
  17. Quantify on Nanodrop with EB as blank.

Recipes

3M Sodium Acetate

Combine in 15mL Falcon tube:

  • 1.23g Sodium Acetate (m.w. = 82.03g/mol)
  • 5mL ddH2O

Shake to mix. Store in refrigerator.