title: Comparison of DNA isolation using salting-out procedure and automated isolation (MagNA system).
abstract: Isolation of genomic DNA is a key step in genetic analysis. The aim of the study was to evaluate the suitability of isolation of DNA from peripheral blood with manual salting-out procedure and automated MagNA system under specific conditions. The impact of storage conditions, type of material (whole blood or blood cells), and method used for DNA extraction were evaluated in terms of DNA yield, its purity, and integrity. Fresh material, and material stored at 2-8°C for 1-4 weeks and frozen at -80°C were tested. For fresh samples, salting-out method gives higher yield than MagNA, irrespectively, on material used. Neither the yield of salting-out method nor its purity decreases during the storage of the samples in the fridge (2-8°C) during 4 weeks. Concerning MagNA, storage of blood cells in the fridge decreases the yield of DNA as well as its purity. For frozen samples, for whole blood, MagNA gives better results while for blood cells, salting-out method seems to be better. For fresh samples, salting-out method is the preferred one, and both whole blood and blood cells can be used. For frozen samples, the preferred method depends on the material.
authors.affiliation: a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Marta
authors.initials: M
authors.lastname: Kalousová
authors.affiliation: a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Levová
authors.affiliation: a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Aleš A
authors.initials: AA
authors.lastname: Kuběna
authors.affiliation: a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Jáchymová
authors.affiliation: b Institute of Inherited Metabolic Disorders, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Věra
authors.initials: V
authors.lastname: Franková
authors.affiliation: a Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine , Charles University and General University Hospital , Prague , Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Zima
mesh_terms: D000328:Adult; D001793:Blood Preservation; D005591:Chemical Fractionation / Q000379:methods; D004247:DNA / Q000097:blood* / Q000302:isolation & purification*; D005260:Female; D005615:Freezing; D006801:Humans; D008297:Male; D008875:Middle Aged; D012492:Salts / Q000737:chemistry*
chemical_list: D012492:Salts; D004247:DNA
keywords: Biobank; DNA isolation; MagNA; blood cells; salting out; whole blood