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Protocol for GenomePlex Whole Genome Amplification from Plant

BeadArray analysis.

II. Product Components

  • Sigma GenElute Plant Genomic DNA Miniprep (G2N10)
  • GenomePlex Whole Genome Amplification Kit (WGA1)
  • GenElute PCR Clean-Up Kit (NA1020)

III. Materials to be Supplied by the User

  • Plant samples
  • 65 C water bath or heating block
  • 1.5 ml microcentrifuge tubes for lysis, Product Code T9661
  • Microcentrifuge (with rotor for 2ml tubes)
  • Ethanol (absolute), Product Code E7023
  • Molecular Biology Reagent Water, Product Code W4502
  • JumpStart Taq DNA Polymerase (D9307)

IV. Protocol for Extraction of DNA from Plant

  • Performed with Sigma GenElute Plant Genomic DNA Miniprep (G2N10)
    1. Grind approximately 50 mg leaf punch into a fine powder with liquid nitrogen. Keep the sample on ice for immediate use or freeze at -70 C.
    2. Add 350 l of Lysis Solution (Part A) and 50 l of Lysis Solution (Part B) and thoroughly mix by vortexing. A white precipitate will form upon the addition of Lysis Solution Part B.
    3. Incubate the mixture at 65 C for 10 minutes with occasional inversion to dissolve the precipitate.
    4. Add 130 l of Precipitation Solution, mix by inversion, and place the sample on ice for 5 minutes.
    5. Centrifuge at maximum speed (12,000 to 16,000 x g) for 5 minutes to pellet the cellular debris, proteins and polysaccharides.
    6. Carefully pipette the supernatant onto a GenElute Filtration Column (blue insert with a 2 ml collection tube).
    7. Centrifuge at maximum speed for 1 minute. Discard the Filtration Column and retain the collection tube.
    8. Add 700 l of Binding Solution directly to the flow-through (liquid from step 7). Mix thoroughly by inversion.
    9. Insert GenElute Miniprep Binding Column (red o-ring) into the provided microcentrifuge tube.
    10. Add 500 l of the
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