Whole Genome Amplication

Purification Protocol

Purification of amplified products performed withGenElute™ PCR Clean-up Kit

(Product No. NA1020)

  1. Insert a GenElute Miniprep Binding Column (with a blue o-ring) into a provided collection tube.
  2. Add 0.5 ml of the Column Preparation Solution to each Miniprep column and centrifuge at 12,000 × g for 30 seconds to 1 minute. Discard the eluate.
    Note: The Column Preparation Solution maximizes binding of the DNA to the membrane resulting in more consistent yields.
  3. Add 5 volumes of Binding Solution to 1 volume of the PCR reaction and mix.
    Example: add 500 µl of Binding Solution to 100 µl of the PCR reaction.
  4. Transfer the solution into the binding column. Centrifuge the column at maximum speed (12,000–16,000 × g) for 1 minute. Discard the eluate, but retain the collection tube.
  5. Replace the binding column in the collection tube. Apply 0.5 ml of diluted Wash Solution to the column and centrifuge at maximum speed for 1 minute. Discard the eluate, but retain the collection tube.
    Note: Be sure to add ethanol to the Wash Solution Concentrate prior to first time use. See Preparation Instructions.
  6. Replace the column in the collection tube. Centrifuge the column at maximum speed for 2 minutes, without any additional wash solution, to remove excess ethanol. Discard any residual eluate as well as the collection tube.
  7. Transfer the column to a fresh 2 ml collection tube. Apply 50 µl of Elution Solution or water to the center of each column. Incubate at room temperature for 1 minute.
    Note: When eluting with water, make sure that the pH of the water is between 5.5 and 8.5. Elution may also be performed using the Elution Solution diluted 10-fold with water.
  8. To elute the DNA, centrifuge the column at maximum speed for 1 minute. The PCR amplification product is now present in the eluate and is ready for immediate use or storage at –20 °C .