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    • 7. 发明申请
    • METHODS FOR DETECTING SMALL RNA SPECIES
    • 检测小RNA物种的方法
    • WO2008119072A1
    • 2008-10-02
    • PCT/US2008/058758
    • 2008-03-28
    • ILLUMINA, INC.FAN, Jian-bingCHEN, Jing
    • FAN, Jian-bingCHEN, Jing
    • C12Q1/68
    • C12Q1/6844C12Q2525/207C12Q2525/173C12Q2521/107C12Q2525/191
    • The invention provides a method of detecting small target nucleotide sequences, in particular, small RNA species that are present in a sample. The method generally comprises a poly- A polymerization step or a ligation step to add a universal sequence to the 3 '-end of all RNA molecules, followed by a universal primer- mediated cDNA synthesis, solid-phase selection, assay oligo annealing, extension and PCR amplification/labeling. The method of the invention can be practiced to amplify and label a small amount of miRNA or other ncRNA. The resulting amplification product can be read out on a universal array or an array with miRNA-specific or ncRNA- specific probes. The invention has multiple embodiments, including methods, compositions, and kits. In general, the nucleic acids, compositions, and kits comprise materials that are useful in carrying out the methods of the invention or are produced by the methods, and that can be used to detect small target nucleic acid sequences present in samples, in particular, small RNA species.
    • 本发明提供了检测小目标核苷酸序列,特别是存在于样品中的小RNA种类的方法。 该方法通常包括聚A聚合步骤或连接步骤,以将通用序列添加到所有RNA分子的3'末端,随后通用引物介导的cDNA合成,固相选择,测定寡聚退火,延伸 和PCR扩增/标记。 可以实施本发明的方法来扩增和标记少量的miRNA或其他ncRNA。 可以在通用阵列或具有miRNA特异性或ncRNA特异性探针的阵列上读出所得到的扩增产物。 本发明具有多种实施方案,包括方法,组合物和试剂盒。 通常,核酸,组合物和试剂盒包含可用于实施本发明方法或通过该方法生产的材料,并且可用于检测样品中存在的小靶核酸序列,特别是, 小RNA种类。