Assay for the quantification of small-sized fragmented genomic DNA.
This paper presents a simple assay for the direct quantification of small-sized (up to 1000 bp) fragmented DNA based on the differential separation between large- and small-sized DNA by polyethylene glycol precipitation. The assay can be applied to as low as 1 microg ml(-1) DNA and has a very low DNA detection limit (0.01 microg ml(-1) fragmented DNA), which can be further lowered at least 10-fold with anion exchange chromatography. The assay quantifies for the first time a more direct indicator of the extent of DNA damage than the usually assessed DNA nicks and base modifications, since small-sized fragmented DNA represents an unrepairable DNA damage of necrotic and apoptotic events that are related to normal and abnormal biological conditions.
Georgiou
Christos D
CD
Section of Genetics, Cell Biology and Development, Department of Biology, University of Patras, 26100 Patras, Greece. c.georgiou@upatras.gr
Patsoukis
Nikolaos
N
Papapostolou
Ioannis
I
D006690:Bisbenzimidazole; D005591:Chemical Fractionation / Q000379:methods; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange / Q000379:methods; D004247:DNA / Q000032:analysis*; D053938:DNA Fragmentation; D004587:Electrophoresis, Agar Gel; D005456:Fluorescent Dyes; D016678:Genome; D008970:Molecular Weight; D011092:Polyethylene Glycols; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D011092:Polyethylene Glycols; D004247:DNA; D006690:Bisbenzimidazole