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Protocol for GenomePlex Whole Genome Amplification from Blood Card

tide primers and a limited number of cycles. This technology maintains the genetic representation with concordance >99.8% in genotyping results from comparing genomic DNA and GenomePlex amplified DNA1. In addition to SNP genotyping, downstream applications also include performing TaqMan assays and BeadArray analysis.

II. Product Components

  • GenElute Blood Genomic DNA Kit (NA2000)
  • GenomePlex Whole Genome Amplification Kit (WGA1)
  • GenElute PCR Clean-Up Kit (NA1020)
III. Materials to be Supplied by the User
  • Blood Card
  • 1.5 ml microcentrifuge tubes for lysis
  • Microcentrifuge (with rotor for 2ml tubes)
  • Ethanol (absolute) Product Code E7023
  • Molecular Biology Reagent Water, Product Code W4502
  • JumpStart Taq DNA Polymerase, Product Code D9307
  • 55 C water bath or heating block
IV. Protocol for Extraction of DNA from Blood Card
  • Performed with GenElute Blood Genomic DNA (NA2000)
    1. Cut a disc from a dried blood card (200 l spotted) into several 2 mm by 2 mm pieces and place the pieces into a 1.5 ml microcentrifuge tube.
    2. Add 40 l Proteinase K and 1.0 ml Resuspension Solution.
    3. Add 300 l of Lysis Solution C and vortex thoroughly for 15 seconds.
    4. Incubate at 55 C for 10 minutes.
    5. After the incubation, transfer the liquid (discard blood card remaining in the microcentrifuge tube) to a 15ml conical tube.
    6. Add 500 l of Column Preparation Solution to the GenElute Miniprep Binding Column (red o-ring) and centrifuge at 12,000 x g for 1 minute. Discard the flow-through liquid.
    7. Add 900 l of 95-100% ethanol to the lysate in the 15 ml conical tube and mix thoroughly by vortexing 5 to 10 seconds.
    8. Transfer the contents of the tube into the treated column from step 4. Centrifuge at ≥ 6,500 x g for
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