Phenol extraction revisited: a rapid method for the isolation and preservation of human genomic DNA from whole blood.
We report a fast and simple DNA isolation method from whole blood. It avoids cell separation and lysis steps and consists of three successive solvent extractions and an ethanol precipitation. All the steps are carried out at room temperature. The main advantage of this method is the immediate sample inactivation achieved by mixing the blood sample with Tris-HCl (pH 8.0) saturated phenol, thus minimizing the biohazard involved in the subsequent manipulation of the samples potentially contaminated with infectious agents (the procedure has been called SP for 'straight phenol'). In addition, extensive field sample collections are facilitated by the fact that the SP procedure can be stopped right after the simple manipulation of mixing the blood sample with the phenol; neither freezing nor refrigeration of the sample proved to be required. At this stage, the nucleases as well as infectious agent are inactivated and the rest of the protocol can wait to be carried out in the laboratory. In fact, the DNA preparation can be resumed after prolonged storage of the blood-phenol mix (up to 72 days has been checked in our laboratory) at room temperature without affecting the yield. The SP protocol may be scaled up, when large quantities of DNA are needed, or scaled down to smaller volumes, such as fingerprick blood samples.
Albariño
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CG
Instituto de Bioquímica y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de la Plata, Argentina.
Romanowski
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V
D001483:Base Sequence; D001793:Blood Preservation; D004247:DNA / Q000097:blood*; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D010636:Phenols / Q000737:chemistry*; D016133:Polymerase Chain Reaction
D010636:Phenols; D004247:DNA