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Protocol for GenomePlex Whole Genome Amplification ,,,from Buccal Swab

rdance >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 Mammalian Genomic DNA Miniprep Kit (G1N10)
  • GenomePlex WGA kit (WGA1)
  • GenElute PCR Clean-Up Kit (NA1020)

III. Materials to be Supplied by the User

  • Buccal swab
  • 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
  • 70 C water bath or heating block

IV. Protocol for Extraction of DNA from Buccal Swab

  • Performed with GenElute Mammalian Genomic DNA Miniprep (G1N10)
    1. Collected swabs were dried at room temperature for 15 minutes.
    2. Add 280 l of Lysis Solution T and 20 l of Proteinase K. Insert the swab and gently spin.
    3. Cap the tube and mix by vortexing.
    4. Incubate the sample at 55 C for 20 minutes with occasional vortexing.
    5. Add 200 l of Lysis Solution C and vortex thoroughly for 15 seconds.
    6. Incubate at 70 C for 10 minutes.
    7. Add 500 l of Column Preparation Solution to each GenElute Miniprep Binding Column (red o-ring) and centrifuge at 12,000 X g for 1 minute. Discard the flow-though liquid.
    8. Add 200 l of 95-100% ethanol to the lysate from step 6.
    9. Mix thoroughly by vortexing and add the entire contents of the tube into the binding column.
    10. Centrifuge at ≥ 6,500 X g for 1 minute.
    11. Discard the collection tube containing the flow through and place the binding column in a new 2 ml collection t
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