Zheng Ke-Wei, Zhang Jia-Yu, Tan Zheng
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.
Methods Mol Biol. 2019;2035:243-255. doi: 10.1007/978-1-4939-9666-7_14.
In vitro transcription of double-stranded DNA (dsDNA) induces a formation of two types of G-quadruplexes, intramolecular DNA G-quadruplexes or DNA:RNA hybrid G-quadruplexes, in clusters of guanine tracts. The formation of G-quadruplexes can be characterized by native PEG polyacrylamide gel electrophoresis, DMS footprinting, and ligand-induced photocleavage footprinting. The type of G-quadruplexes can be distinguished by transcription with 7-deaza-GTP (dzGTP) or 4-thio-uridine-5'-triphosphate (4U) to detect the participation of RNA transcript.
双链DNA(dsDNA)的体外转录会在鸟嘌呤序列簇中诱导形成两种类型的G-四链体,即分子内DNA G-四链体或DNA:RNA杂交G-四链体。G-四链体的形成可以通过非变性聚乙二醇聚丙烯酰胺凝胶电泳、二甲基亚砜足迹法和配体诱导的光裂解足迹法来表征。G-四链体的类型可以通过用7-脱氮鸟苷三磷酸(dzGTP)或4-硫代尿苷-5'-三磷酸(4U)进行转录来区分,以检测RNA转录本的参与情况。