发明名称 Methods, Reagents and Kits for Detection of Nucleic Acid Molecules
摘要 Methods, reagents and kits are provided for the production and use in detection assays of labeled nucleic acid molecules wherein a labeling molecule is attached directly to the 3′ end of the nucleic acid molecules.
申请公布号 US2014296092(A1) 申请公布日期 2014.10.02
申请号 US201414187806 申请日期 2014.02.24
申请人 Genisphere, LLC 发明人 Getts Robert C.;Kadushin James;Bowers Jessica
分类号 C12Q1/68 主分类号 C12Q1/68
代理机构 代理人
主权项 1. A method for detecting an antisense probe on a solid support comprising: a) contacting a solid support having thereon an antisense probe comprising the complementary nucleotide sequence of a target nucleic acid molecule with a labeled target nucleic acid molecule comprising a linear dendritic polynucleotide, the linear dendritic polynucleotide comprising an oligonucleotide extension sequence having a 5′ phosphate group, and first, second and third polynucleotide monomers bonded together by hybridization in a 5′-3′ orientation, each polynucleotide monomer, prior to being hybridization bonded to one another, having first, second and third single stranded hybridization regions; the third single stranded hybridization region of the first polynucleotide monomer being hybridization bonded to the first single stranded hybridization region of the second polynucleotide monomer, and the third single stranded hybridization region of the second polynucleotide monomer being hybridization bonded to the first single stranded hybridization region of the third polynucleotide monomer, and the first single stranded region of the first polynucleotide monomer being hybridization bonded to a bridging oligonucleotide, wherein the second single stranded hybridization regions of the linear dendritic polynucleotide are hybridization bonded to one or more labeled oligonucleotides comprising one or more label molecules; and wherein the bridging oligonucleotide is hybridization bonded to the target nucleic acid molecule and the oligonucleotide extension sequence. b) incubating the solid support and the labeled target nucleic acid molecule for a time and at a temperature sufficient to enable the labeled target nucleic acid molecule to hybridize to the antisense probe; c) washing the solid support to remove unhybridized labeled target nucleic acid molecule; and d) detecting the signal from the hybridized labeled target nucleic acid molecule, thereby detecting the antisense probe on the solid support.
地址 Hatfield PA US