Líffræðifélag Íslands
Líffræðiráðstefnan 2015
Erindi/veggspjald / Talk/poster E43
Bjarki Guðmundsson (1,2,3), Hans G. Þormar (1,2), Albert Sigurðsson (1), Wendy Dankers (1), Margrét Steinarsdóttir (3), Davíð Ólafsson (1,4), Anna M. Halldórsdóttir (4), Jón J. Jónsson (1,3)
1. Lífefna- og sameindalíffræðistofa læknadeildar, Háskóli Íslands, 2. Lífeind ehf., 3. Erfða- og sameindalæknisfræðideild, Landspítali, 4. Blóðbankinn, Landspítali, Reykjavík.
Kynnir / Presenter: Bjarki Guðmundsson
Tengiliður / Corresponding author: Bjarki Guðmundsson (bjarkigu@hi.is)
Detection of DNA damage has many applications including testing quality of DNA samples in biobanks, monitoring complex molecular procedures, genotoxicity testing, diagnosing genome instability, and in cancer theragnostics. We tested the suitability of the Northern Lights Assay (NLA) for these applications. NLA is based on Two-Dimensional Strandness-Dependent Electrophoresis (2D-SDE), a technique of nucleic acid separation based on size, standness, and conformation changes induced by damage. Each specimen is analyzed in sample pairs of non-digested DNA to detect single- and double-stranded breaks and MboI-digested DNA to detect other lesions. We tested NLA on various samples including DNA in solution, intermediate products of complex procedures, various cell cultures treated with genotoxic agents, and body fluid samples from patients. NLA could detect single-stranded breaks, double-stranded breaks, interstrand and intrastrand DNA crosslinks, single-stranded DNA, bulky lesions and mismatches in all types of samples. The sensitivity of NLA was sufficient for DNA damage detection at biologically relevant concentrations of damaging agents. The main advantage of NLA compared to the comet assay was detection of both intrastrand and interstrand crosslinks and other abnormal DNA in a direct manner. In addition, NLA has the ability to analyze DNA damage in cell-free systems. NLA is a versatile method for comprehensive and simultaneous analysis of various types of DNA damage both in purified DNA and in biological samples from cells and body fluids. NLA is useful in biomedical research and diagnostics.