Isolation of cell-free DNA from plasma by chromatography on short monolithic columns and quantification of non-apoptotic fragments by real-time polymerase chain reaction (CROSBI ID 148647)
Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija
Podaci o odgovornosti
Ivančić-Jelečki, Jelena ; Brgles, Marija ; Šantak, Maja ; Forčić, Dubravko
engleski
Isolation of cell-free DNA from plasma by chromatography on short monolithic columns and quantification of non-apoptotic fragments by real-time polymerase chain reaction
Human plasma is an important medical substance and a raw material for production of various therapeutics. During blood sampling, storage and processing, genomic DNA is released into plasma from nucleated blood cells that are damaged in the course of the procedure. In order to determine the concentration of contaminating DNA in plasma, we developed a method for DNA isolation by using anion-exachange chromatography on a BIA Separations CIM (convective interaction media) diethylaminoethyl column. DNA was quantified by SYBR Green based real-time polymerase chain reaction. The concentration of cell-free, non-apoptotic DNA in plasma ranged between 0.06 and 22.5 ng/ml. As substantial volumes of plasma or whole blood are administered directly into the vascular system, a recipient is exposed to high amounts of cell-free DNA, several orders of magnitude higher than the amount found in other biologicals.
Cell-free DNA; DNA isolation; Chromatography; Plasma; Convective Interaction Media (CIM); Quantitative PCR
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Podaci o izdanju
1216 (13)
2009.
2717-2724
objavljeno
0021-9673
10.1016/j.chroma.2008.10.087
Povezanost rada
Kemija, Temeljne medicinske znanosti, Biologija