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Protocol for GenomePlex Whole Genome Amplification ,,,from Whole Blood

brary is then PCR amplified using universal oligonucleotide 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 WGA kit (WGA1)
  • GenElute PCR Clean-Up Kit (NA1020)
III. Materials to be Supplied by the User
  • Whole Blood
  • 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 Whole Blood
  • Performed with GenElute Blood Genomic DNA (NA2000)
    1. Place 20 l of Proteinase K into a 1.5 ml microcentrifuge tube and add 200 l of whole blood to the tube.
    2. Add 200 l of Lysis Solution C and vortex thoroughly for 15 seconds.
    3. Incubate at 55 C for 10 minutes.
    4. 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-thorough liquid.
    5. Add 200 l of 95-100% ethanol to the lysate from step 3 and mix thoroughly by vortexing 5 to 10 seconds.
    6. Transfer the entire contents of the tube into the treated column (from step 4). Centrifuge at ≥ 6,500 x g for 1 minute.
    7. Discard the collection tube and flow through and place the column in a new 2 ml collection tube.
    8. Add 500 l of Prewash Solution (be sure to dilute with ethanol prior to first u
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