Pre-Amplification Protocol for PicoPLEX WGA Kit

Protocol

  1. Combine Extraction Cocktail components and mix well:
    EXTRACTION COCKTAIL VOLUME PER 5 SAMPLES
    Extraction Enzyme Dilution Buffer 24 μl
    Cell Extraction Enzyme 1 μl
    Total volume 25 μl
  2. Add 5 μl of freshly-prepared Extraction Cocktail to each 5 μl cell sample or control DNA sample prepared by the sample preparation methods.

  3. Incubate sample in a thermal cycler as follows:
    CYCLES
    TEMP
    TIME
    1
    75°C
    10 minutes
    95°C
    4 minutes
    12°C
    Hold
  4. Combine Pre-Amp Cocktail components and mix well:
    PRE-AMP COCKTAIL VOLUME PER 5 SAMPLES
    Pre-Amp Reaction Mix 24 μl
    Pre-Amp Enzyme 1 μl
    Total volume 25 μl
  5. Add 5 μl of Pre-Amp Cocktail to each cell or control sample.

  6. Incubate sample according to the thermal cycler program below:
    CYCLES TEMP
    TIME
    1 95°C
    2 minutes
    12 95°C 15 seconds
    15°C 50 seconds
    25°C 40 seconds
    35°C 30 seconds
    65°C 40 seconds
    75°C 40 seconds
    1 4°C Hold
  7. Briefly centrifuge sample and place Pre-Amp incubation product on ice.