An efficient and simple method of DNA extraction from whole blood and cell lines to identify infectious agents

J Virol Methods. 1991 Sep;34(1):105-12. doi: 10.1016/0166-0934(91)90126-k.

Abstract

Routine methods of extraction of DNA from blood involve the enrichment of cells by Ficoll-Hypaque gradient centrifugation followed by lysis of the cells with extraction buffer, proteinase K digestion of the lysate, and phenol:chloroform-isoamyl alcohol extraction. These methods generally require large amounts of blood, which poses a problem with pediatric patients. To overcome this, we developed a new method of extracting DNA directly from whole blood. This method involves the treatment of whole blood with an equal volume of NaI (3 M final concentration) followed by chloroform:isoamyl alcohol extraction to clear hemoglobin and cell debris. The clear aqueous layer is then mixed with isopropanol to obtain DNA. A large number of samples can easily be handled by this extraction procedure, as it can be carried out in 30 min and requires only a microcentrifuge.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • Cell Line
  • Centrifugation, Density Gradient
  • DNA, Viral / blood
  • DNA, Viral / isolation & purification*
  • HIV / genetics
  • HIV / isolation & purification
  • HIV Infections / microbiology
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • Sodium Iodide

Substances

  • DNA, Viral
  • Sodium Iodide