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Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood ,,, Mini Kit

  • Centrifuge vial at 1,500 x g for 10 minutes in a clinical centrifuge.
  • Pipette 200 l of buffy coat for each sample to be processed into a fresh microcentrifuge tube.
    The buffy coat is seen as a milky white layer between the clear upper plasma layer and the lower red blood cell layer. The buffy coat layer will be very viscous. Take care when pipetting. Pipetting the buffy coat may be easier if some of the clear plasma layer is removed from the top first.
  • Add 20 l of Proteinase K solution to the microcentrifuge tube. Add 350 l Solution G1 to the same tube.
  • Vortex vigorously for 5 seconds. This step is very important due to the presence of high numbers of white blood cells.
  • Incubate samples in a Thermomixer at 70C, 900 rpm, for 10 minutes. A water bath may be used with intermittent vortexing of the samples if a Thermomixer is not available.
  • Add 200 l Solution G2 to the tube. Vortex vigorously for 5 seconds. Place a spin column in a fresh microcentrifuge tube. Transfer the sample to the spin column assembly by pouring or pipetting. Incubate the sample at room temperature for 1 minute.
  • Centrifuge the sample for 2 minutes at 12,00016,000 x g. Remove the spin column and decant flow-through. Place the spin column back into the same tube.
  • Add 600 l Diluted Wash Buffer to the spin column. Centrifuge for 1 minute at 12,00016,000 x g. Remove the spin column and decant flow-through. Place the spin column back into the same tube.
  • Add 400 l Diluted Wash Buffer to the spin column. Centrifuge for 3 minutes at
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