title: An efficient and simple method of DNA extraction from whole blood and cell lines to identify infectious agents.
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.
authors.affiliation: Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
authors.forename: V N
authors.initials: VN
authors.lastname: Loparev
authors.forename: M A
authors.initials: MA
authors.lastname: Cartas
authors.forename: C E
authors.initials: CE
authors.lastname: Monken
authors.forename: A
authors.initials: A
authors.lastname: Velpandi
authors.forename: A
authors.initials: A
authors.lastname: Srinivasan
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification*; D006678:HIV / Q000235:genetics / Q000302:isolation & purification; D015658:HIV Infections / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012974:Sodium Iodide
chemical_list: D004279:DNA, Viral; D012974:Sodium Iodide