Genomic DNA extraction by Salting-out

ABELLEYRO, MIGUEL MARTÍN; RADIC CLAUDIA PAMELA; MARCHIONE, VANINA DANIELA; DE BRASI, CARLOS DANIEL; ROSSETTI, LILIANA CARMEN

World Federation for Hemophilia Genetics Laboratory manual.

World Federation of Hemophilia

Año: 2022;

The quality/quantity of the genomic DNA sample (gDNA) is the principal factor to succeed in achieving most types of genetic tests. Laboratories for Genetic Testing in Hemophilia do not evade this point being vital to count with a high-quality gDNA substrate from patients and controls. A proper patient?s gDNA should show high molecular weight (>25 kb in average) and purity (free of organic solvents and low amounts of RNA and proteins). These features are particularly important for hemophilia molecular testing in which the characterization of structural variants such as those causing about a half of severe hemophilia A (HA) patients is routine. For most gene testing applications including the screening and characterization of the pathogenic variant associated with hemophilia, a gDNA sample extracted from peripheral blood leukocytes is acceptable. Peripheral blood (PB) samples should be maintained at approximately 10-15° C prior use following the rules of biosafety and biosecurity for traveling inside and outside health care institutions. It is crucial to indicate that PB samples should all be treated as potential sources of infection (particularly Hepatitis C, Hepatitis B an Human Immunodeficiency viruses). Identical to any laboratory procedure, the salting out method for gDNA extraction should take into account the hazards of storage, handling and disposal of the involved materials. These details should be properly addressed in a standard operating protocol (SOP), which should be ideally approved by a local Institutional Biosecurity Committee. The Laboratory Head or Director is responsible for following the rules of good practice and biosafety by all the personnel involved in the Laboratory. The salting out method for gDNA extraction presented here was adapted from those described by Lahiri & Nuremberg (1).