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Protocol for GenomePlex Whole Genome Amplification from Plant

Column Preparation Solution to each Miniprep Column and centrifuge at 12,000 x g for 1 minute. Discard the flow-through liquid.
  • Pipette 700 l flow-through (from step 8) onto the Miniprep Column prepared in the previous step .
  • Centrifuge at maximum speed for 1 min and discard the flow-through.
  • Apply the remaining lysate (from step 8) and repeat centrifugation for 1 minute at maximum speed and discard the flow-through.
  • Place the Binding Column in a fresh 2 ml collection tube and apply 500 l diluted Wash Solution to the column (be sure to add ethanol to Wash Solution Concentrate prior to first time use).
  • Centrifuge at maximum speed for 1 minute and discard flow through and retain the collection tube.
  • Add another 500 l of diluted Wash Solution to the column and centrifuge at maximum speed for 3 minutes to dry the column.
  • Transfer the binding column to a fresh 2 ml collection tube.
  • Apply 100 l of pre-warmed (65 C) Elution Solution to the column and centrifuge at maximum speed for 1 minute.
  • Repeat the elution.
  • Store the eluted DNA at -20 C.

    V. Protocol for GenomePlex Whole Genome Amplification from Plant

    • Performed with GenomePlex Whole Genome Amplification Kit (WGA1)

      Fragmentation

      1. Prepare DNA solution of 1 ng/l from plant genomic DNA extraction 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 t
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