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    • 45. 发明授权
    • Nucleic acid sequencing
    • 核酸测序
    • US07094531B1
    • 2006-08-22
    • US09341641
    • 1998-01-15
    • Gunter SchmidtAndrew Hugin Thompson
    • Gunter SchmidtAndrew Hugin Thompson
    • C12Q1/68C12N15/00C12N15/63C12N1/20C07H21/04
    • C12Q1/6874C12Q1/6872
    • Provided is a method for sequencing DNA, which comprises: (a) obtaining a target DNA population comprising one or more single-stranded DNAs to be sequenced, each of which is present in a unique amount and bears a primer to provide a double-stranded portion of the DNA for ligation thereto; (b) contacting the DNA population with an array of hybridisation probes, each probe comprising a label cleavably attached to a known base sequence of predetermined length, the array containing all possible base sequences of that predetermined length; (c) removing all unligated probes; followed by the steps of: (d) cleaving the ligated probes to release each label; (e) recording the quantity of each label; and (f) activating the extended double-stranded portion to enable ligation thereto; wherein (g) steps (b) to (f) are repeated in a cycle for a sufficient number of times to determine the sequence of the or each single-stranded DNA by determining the sequence of release of each label.
    • 提供了一种用于测序DNA的方法,其包括:(a)获得包含待测序的一个或多个单链DNA的靶DNA群体,其中每一个以独特的量存在并承载引物以提供双链 DNA连接部分; (b)使DNA群体与杂交探针阵列接触,每个探针包含与预定长度的已知碱基序列可切割地连接的标记,该阵列包含该预定长度的所有可能的碱基序列; (c)清除所有未连接的探针; 其次是以下步骤:(d)切割连接的探针以释放每个标签; (e)记录每个标签的数量; 和(f)活化延伸的双链部分以使其能够连接; 其中(g)步骤(b)至(f)在循环中重复足够多次,以通过确定每个标记的释放顺序来确定该单链DNA或每个单链DNA的序列。
    • 50. 发明授权
    • Chimeric oligonucleotides and uses thereof in the identification of antisense binding sites
    • 嵌合寡核苷酸及其在鉴定反义结合位点中的用途
    • US06355418B1
    • 2002-03-12
    • US09043149
    • 1998-03-13
    • Gunter Schmidt
    • Gunter Schmidt
    • C12Q168
    • C07H21/00
    • A chimaeric oligonucleotide library for use in identifying an antisense binding site in a target mRNA, comprising a plurality of distinct chimaeric oligonucleotides capable of hybridizing to mRNA to form a duplex, the nucleotide sequences of which each have a common length of 7 to 20 bases and are generated randomly or generated from information characterizing the sequence of the target mRNA, wherein substantially all the nucleotide sequences of said common length which are present as sub-sequences in the target mRNA are present in the library, and wherein each nucleotide sequence comprises: a) a recognition region comprising a sequence of nucleotides which is recognizable by a duplex-cutting RNAase when hybridized to the mRNA, and b) a flanking region comprising a sequence of chemically-modified nucleotides which binds to the mRNA sufficiently tightly to stabilize the duplex for cutting of the mRNA in the duplex by the duplex-cutting RNAase, wherein the nucleotides constituting the flanking region are different from those constituting the recognition region, and wherein each oligonucleotide is protected against exonuclease attack.
    • 用于鉴定靶mRNA中的反义结合位点的嵌合寡核苷酸文库,其包含能够与mRNA杂交以形成双链体的多个不同的嵌合寡核苷酸,其核苷酸序列的长度为7至20个碱基, 由表达靶mRNA序列的信息产生,其中基本上所有作为目标mRNA中的亚序列存在的所述共同长度的所有核苷酸序列存在于文库中,并且其中每个核苷酸序列包含: )识别区域,其包含当与mRNA杂交时由双链切割RNA酶识别的核苷酸序列,和b)包含化学修饰的核苷酸序列的侧翼区域,所述化学修饰的核苷酸序列足够紧密地结合到mRNA上以稳定双链体 通过双链切割RNA酶切割双链体中的mRNA,其中构成fl 贴附区域与构成识别区域的区域不同,并且其中每个寡核苷酸被保护免受外切核酸酶攻击。