title: A RCA-based assay for analyzing individual strand break in DNA heteroduplex cleavage by restriction endonucleases.
abstract: We have developed a rapid and high-throughput assay based on rolling circle amplification, to distinguish individual strand cleavage of DNA duplexes by restriction endonucleases. As an illustration, we analyzed nicking activity of Nb.BbvCI and uneven cleavage of LNA modified DNA by EcoRI. This assay has potential for analyzing protein-DNA interactions.
authors.affiliation: Key Laboratory of Medical Cell Biology (Ministry of Education), Department of Biochemistry and Molecular Biology, China Medical University, Shenyang, Liaoning 110001, China. yfguan@mail.cmu.edu.cn.
authors.forename: Guojie
authors.initials: G
authors.lastname: Zhao
authors.forename: Tianyu
authors.initials: T
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Hua
authors.initials: H
authors.lastname: Wei
authors.forename: Hong
authors.initials: H
authors.lastname: Shang
authors.forename: Yifu
authors.initials: Y
authors.lastname: Guan
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D053960:DNA Breaks; D004277:DNA, Single-Stranded / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis / Q000235:genetics / Q000378:metabolism
chemical_list: D004277:DNA, Single-Stranded; D009841:Oligonucleotides; C477371:locked nucleic acid; D004247:DNA; D015246:Deoxyribonuclease EcoRI; D003850:Deoxyribonuclease I