TY - JOUR
T1 - Northern lights assay
T2 - A versatile method for comprehensive detection of DNA damage
AU - Gudmundsson, Bjarki
AU - Thormar, Hans G.
AU - Sigurdsson, Albert
AU - Dankers, Wendy
AU - Steinarsdottir, Margret
AU - Hermanowicz, Stefan
AU - Sigurdsson, Stefan
AU - Olafsson, David
AU - Halldorsdottir, Anna M.
AU - Meyn, Stephen
AU - Jonsson, Jon J.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - DNA damage assays have various limitations in types of lesions detected, sensitivity, specificity and samples that can be analyzed. The Northern Lights Assay (NLA) is based on 2D Strandness-Dependent Electrophoresis (2D-SDE), a technique that separates nucleic acids based on length, strandness, structure and conformation changes induced by damage. NLA is run on a microgel platform in 20- 25 min. Each specimen is analyzed in pairs of nondigested DNA to detect single- and double-stranded breaks (DSBs) and Mbo I-digested DNA to detect other lesions. We used NLA to evaluate DNA in solution and isolated from human cells treated with various genotoxic agents. NLA detected and distinguished between single- and DSBs, interstrand and intrastrand DNA crosslinks, and denatured singlestranded DNA. NLA was sufficiently sensitive to detect biologically relevant amount of DNA damage. NLA is a versatile, sensitive and simple method for comprehensive and simultaneous analysis of multiple types of damage, both in purified DNA and in DNA isolated from cells and body fluids. NLA can be used to evaluate DNA quality in biosamples, monitor complex molecular procedures, assess genotoxicity, diagnose genome instability, facilitate cancer theranostics and in basic nucleic acids research.
AB - DNA damage assays have various limitations in types of lesions detected, sensitivity, specificity and samples that can be analyzed. The Northern Lights Assay (NLA) is based on 2D Strandness-Dependent Electrophoresis (2D-SDE), a technique that separates nucleic acids based on length, strandness, structure and conformation changes induced by damage. NLA is run on a microgel platform in 20- 25 min. Each specimen is analyzed in pairs of nondigested DNA to detect single- and double-stranded breaks (DSBs) and Mbo I-digested DNA to detect other lesions. We used NLA to evaluate DNA in solution and isolated from human cells treated with various genotoxic agents. NLA detected and distinguished between single- and DSBs, interstrand and intrastrand DNA crosslinks, and denatured singlestranded DNA. NLA was sufficiently sensitive to detect biologically relevant amount of DNA damage. NLA is a versatile, sensitive and simple method for comprehensive and simultaneous analysis of multiple types of damage, both in purified DNA and in DNA isolated from cells and body fluids. NLA can be used to evaluate DNA quality in biosamples, monitor complex molecular procedures, assess genotoxicity, diagnose genome instability, facilitate cancer theranostics and in basic nucleic acids research.
UR - http://www.scopus.com/inward/record.url?scp=85056588963&partnerID=8YFLogxK
U2 - 10.1093/nar/gky645
DO - 10.1093/nar/gky645
M3 - Article
C2 - 30053193
AN - SCOPUS:85056588963
SN - 0305-1048
VL - 46
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 20
M1 - e118
ER -
