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

se) and centrifuge at ≥ 6,500 x g for 1 minute.
  • Discard the collection tube containing the flow-through and place the column in a new 2 ml collection tube.
  • Add 500 l of Wash Solution (be sure to dilute with ethanol prior to first use) to the binding column and centrifuge at maximum speed (12,000 x g to 16,000 x g) for 3 minutes to dry the binding column.
  • Pipette 200 l of Elution Solution onto the column and centrifuge for 1 minute at ≥ 6,500 x g to elute the DNA.
  • Store the eluted DNA at -20 C or proceed with the next step. V. Protocol for GenomePlex Whole Genome Amplification from Whole Blood
    • Performed with GenomePlex Whole Genome Amplification Kit (WGA1)

      Fragmentation

      1. Prepare DNA solution of 1 ng/l from whole blood extraction protocol described above.
      2. Add 1 l of 10X Fragmentation Buffer to 10 l DNA (1 ng/l) in a PCR tube.
      3. Place the tube in a thermal cycler at 95 C for EXACTLY 4 minutes. Note, the incubation is time sensitive and any deviation may alter results.
      4. Immediately cool the sample on ice and centrifuge briefly.

      Library Preparation

      1. Add 2 l of 1x Library Preparation Buffer.
      2. Add 1 l of Library Stabilization Solution.
      3. Mix thoroughly and place in thermal cycler at 95 C for 2 minutes.
      4. Cool the sample on ice and centrifuge briefly.
      5. Add 1 l Library Preparation Enzyme, mix thoroughly, and centrifuge briefly.
      6. Place sample in thermal cycler and incubate as follows:
        • 16 C for 20 minutes
        • 24 C for 20 minutes
        • 37 C for 20 minutes
        • 75 C for 5 minutes
        • 4 C hold
      7. Remove samples from thermal cycler and centrifuge briefly. Samples may be amplified immediately or stored at - 20 C up to three days.

      Amplification


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