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    • 10. 发明申请
    • NUCLEIC ACID ANALYSIS TECHNIQUES
    • 核酸分析技术
    • WO1997027317A1
    • 1997-07-31
    • PCT/US1997001603
    • 1997-01-22
    • AFFYMETRIX, INC.LOCKHART, David, J.CHEE, MarkGUNDERSON, KevinLAI, ChaoqiangWODICKA, LisaCRONIN, Maureen, T.LEE, DannyTRAN, Huu, M.MATSUZAKI, HajimeMcGALL, Glenn, H.BARONE, Anthony, D.
    • AFFYMETRIX, INC.
    • C12Q01/00
    • C12Q1/6809C07H19/052C07H19/12C07H21/00C12Q1/6816C12Q1/6837C12Q2600/156C40B40/00C12Q2565/501C12Q2521/501
    • The present invention provides a simplified method for identifying differences in nucleic acid abundances (e.g., expression levels) between two or more samples. The methods involve providing an array containing a large number (e.g. greater then 1,000) of arbitrarily selected different oligonucleotide probes where the sequence and location of each different probe is known. Nucleic acid samples (e.g. mRNA) from two or more samples are hybridized to the probe arrays and the pattern of hybridization is detected. Differences in the hybridization patterns between the samples indicates differences in expression of various genes between those samples. This invention also provides a method of end-labeling a nucleic acid. In one embodiment, the method involves providing a nucleic acid, providing a labeled oligonucleotide and then enzymatically ligating the oligonucleotide to the nucleic acid. Thus, for example, where the nucleic acid is an RNA, a labeled oligoribonucleotide can be ligated using an RNA ligase. In another embodiment, the end labeling can be accomplished by providing a nucleic acid, providing labeled nucleoside triphosphates, and attaching the nucleoside triphosphates to the nucleic acid using a terminal transferase.
    • 本发明提供用于鉴定两个或多个样品之间的核酸丰度差异(例如,表达水平)的简化方法。 该方法包括提供包含大量(例如大于1,000个)任意选择的不同寡核苷酸探针的阵列,其中每个不同探针的序列和位置是已知的。 来自两个或更多个样品的核酸样品(例如mRNA)与探针阵列杂交,并检测杂交模式。 样本之间的杂交模式的差异表明这些样品之间各种基因的表达差异。 本发明还提供了一种终止标记核酸的方法。 在一个实施方案中,该方法包括提供核酸,提供标记的寡核苷酸,然后将寡核苷酸酶连接到核酸上。 因此,例如,当核酸是RNA时,可以使用RNA连接酶连接标记的寡核糖核苷酸。 在另一个实施方案中,末端标记可以通过提供核酸,提供标记的核苷三磷酸和使用末端转移酶将核苷三磷酸与核酸连接来实现。