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    • 77. 发明申请
    • POLYMERASE DRIVEN NESA
    • 聚合酶驱动NESA
    • US20140349285A1
    • 2014-11-27
    • US14212124
    • 2014-03-14
    • GEORGETOWN UNIVERSITY
    • Mark DanielsenBerenice AlfonsoBolor Tumurpurev
    • C12Q1/68
    • C12Q1/6844C12Q2521/301C12Q2525/131C12Q2525/307
    • The present invention relates to a novel method for detecting a target polynucleotide having a target sequence, comprising (a) exposing the target polynucleotide to an initiating oligonucleotide; (b) extending the initiating oligonucleotide with an extended sequence complementary to the target sequence; (c) ligating the initiating oligonucleotide sequence with the extended sequence to form a circular oligonucleotide having a nicking endonuclease (NE) recognition/cutting sequence; (d) exposing the circular oligonucleotide to a DNA polymerase and a DNA synthesis primer to synthesize DNA having a NE recognition sequence; (e) exposing the synthesized DNA to a probe having the NE recognition/cutting sequence to form a double stranded DNA having a full NE site; (f) exposing the double stranded DNA to a nicking endonuclease (NE) to cleave the probe; and (g) detecting the cleaved probe. The presence of the cleaved probe indicates the presence of the target polynucleotide.
    • 本发明涉及一种用于检测具有靶序列的靶多核苷酸的新方法,其包括(a)将靶多核苷酸暴露于起始寡核苷酸; (b)以与靶序列互补的延伸序列扩展起始寡核苷酸; (c)将起始寡核苷酸序列与扩展序列连接以形成具有切口核酸内切酶(NE)识别/切割序列的环状寡核苷酸; (d)将环状寡核苷酸暴露于DNA聚合酶和DNA合成引物以合成具有NE识别序列的DNA; (e)将合成的DNA暴露于具有NE识别/切割序列的探针以形成具有完整的NE位点的双链DNA; (f)将双链DNA暴露于切口内切核酸酶(NE)以切割探针; 和(g)检测切割的探针。 切割的探针的存在表明靶多核苷酸的存在。