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Genomic DNA Purification for Blood & Body Fluid

The protocol is used to purify genomic DNA from 100-200 μl whole blood, serum or plasma. Whole blood in fresh, frozen or preserved (such as in EDTA, heparin) forms can be used. If the samples can’t be processed immediately, the samples can be preserved at 4oC or –20oC after addition of lysis buffer.

  1. Add 500 μl Lysis Buffer to the 100 μl whole blood, serum or plasma, mix completely by vortexing 5 sec, incubate for 5 min.

  2. Transfer the lysate to a gDNA binding column. Centrifuge at 11,000 rpm for 30-60 sec. Discard the flowthrough.

  3. Add 400 μl Lysis Buffer to the column, centrifuge at 11,000 rpm for 30-60 sec. Discard the flowthrough.

  4. Add 700 μl DNA Wash Buffer to the column, centrifuge at 11,000 rpm for 30-60 sec. Discard the flowthrough. Make sure that ethanol is added to washing buffer as instructed before use.

  5. Centrifuge additional 30-60 sec at 11,000 rpm. It is critical to remove residual ethanol for optimal elution. Residual ethanol may interfere subsequent applications.

  6. Place the DNA column in a clean 1.5 ml microcentrifuge tube. Add 50-100 µl pre-heated (70oC waterbath) DNA Elution Buffer or Nuclease-Free water to the center of the column, incubate for 1 min, centrifuge at 11,000 rpm for 30-60 sec to elute the DNA.

 

The purified DNA is stable in elution buffer or water if stored at  -20°C or below.

 

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