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    • 13. 发明申请
    • MIRROR BISULFITE ANALYSIS
    • 镜像二硫分析
    • WO2015009844A2
    • 2015-01-22
    • PCT/US2014046876
    • 2014-07-16
    • ZYMO RES CORP
    • JIA XI-YUSUN XUEGUANG
    • G01N33/53
    • C12Q1/6858C12Q2521/531C12Q2523/125C12Q2525/117
    • Reagents and methods for detection and quantitation of DNA hydroxymethylcytosine (5hmC) are provided. Specifically, genome-wide and loci analysis methods are provided that utilizes methylated primers coupled with bisulfite-sequencing for detecting and quantitating 5hmC levels in the genome. This methodology also allows quantitation of 5hmC levels at a single CpG. This method, was used to map and quantitate 5hmC sites of neurons from the human brain. This novel method can determine the exact location and abundance of 5hmC, which with bioinformatic analysis utilizing software and computers can facilitate our understanding of how modified cytosines such as hydroxymethylcytosine play a role in regulating gene expression in the brain.
    • 提供了检测和定量DNA羟甲基胞嘧啶(5hmC)的试剂和方法。 具体而言,提供了使用与亚硫酸氢盐测序偶联的甲基化引物来检测和定量基因组中5hmC水平的全基因组和基因座分析方法。 该方法还可以在单​​个CpG上定量5hmC的水平。 该方法用于绘制和定量人脑中5hmC的神经元位点。 这种新方法可以确定5hmC的确切位置和丰度,利用软件和计算机进行生物信息学分析可以帮助我们理解修饰的胞嘧啶(如羟甲基胞嘧啶)如何在调节大脑基因表达中发挥作用。
    • 15. 发明申请
    • BNA CLAMP METHOD
    • BNA钳法
    • WO2014051076A1
    • 2014-04-03
    • PCT/JP2013076329
    • 2013-09-27
    • BNA INC
    • IMANISHI TAKESHIORITA AYAKOBAN IKUYA
    • C12Q1/68C12N15/09
    • C12Q1/6858C07H19/06C07H19/16C07H21/02C07H21/04C12Q2525/117C12Q2535/131C12Q2549/126
    • Provided is an examination method allowing highly sensitive and highly accurate detection of a target nucleic acid (for example a mutant gene), while being simple and low cost. More specifically, in a nucleic acid amplification reaction, an oligonucleotide analog containing 1 or 2 or more of at least one kind of unit structure of each kind of nucleotide analogs represented by the general formula (I) (symbols in the formula are as defined in the description) or similar, or a salt thereof, is used as clamp nucleic acid, the amplification of a non target nucleic acid (for example a wild-type gene) in a sample is inhibited, and thus a target nucleic acid is selectively amplified and the amplified nucleic acid is detected.
    • 本发明提供了一种检测方法,其能够简单且成本低,能够高精度地检测靶核酸(例如突变型基因)。 更具体地说,在核酸扩增反应中,含有1或2以上的通式(I)表示的各种核苷酸类似物的至少一种单元结构的寡核苷酸类似物(式中的符号如 描述)或类似物或其盐用作钳位核酸,样品中非靶核酸(例如野生型基因)的扩增被抑制,因此靶核酸被选择性扩增 并检测扩增的核酸。
    • 20. 发明申请
    • POLYMERASE INCORPORATION OF NON-STANDARD NUCLEOTIDES
    • 聚合物纳入非标准核素
    • WO2009154733A2
    • 2009-12-23
    • PCT/US2009/003595
    • 2009-06-16
    • BENNER, Steven, AlbertCHEN, FeiYANG, Zunyi
    • BENNER, Steven, AlbertCHEN, FeiYANG, Zunyi
    • C12Q1/68C12P19/40
    • C12Q1/6853C12P19/34C12Q1/686C12Q2521/101C12Q2525/117C12Q2525/191
    • The disclosed invention teaches processes to amplify oligonucleotides by contacting templates and primers with DNA polymerases and triphosphates of non-standard nucleotides, which form nucleobase pairs fitting the standard Watson-Crick geometry, but joined by hydrogen bonding patterns different from those that join standard A:T and G:C pairs. Thus, this invention relates to nucleotide analogs and their derivatives that, when incorporated into DNA and RNA, expand the number of replicatable nucleotides beyond the four found in standard DNA and RNA. The invention further relates to polymerases that incorporate those non-standard nucleotide analogs into oligonucleotide products using the corresponding triphosphate derivatives, and more specifically, polymerases and non-standard nucleoside triphosphates that support the polymerase chain reaction (PCR), including PCR where the products contain more than one non-standard nucleotide unit. Examples are provides that show this process using 6-amino-5-nitro-3-(l'-beta- D-2'-deoxyribofuranosyl)-2(lH)-pyridone to implement the non-standard "small" donor-donor- acceptor (pyDDA) hydrogen bonding pattern, and 2-amino-8-(r-beta-D-2'-deoxyribofuranosyl)- imidazo[l,2-α]-l,3,5-triazin-4(8H)-one to implement the "large" acceptor-acceptor-donor (puAAD) pattern.
    • 所公开的发明教导了通过使模板和引物与DNA聚合酶和非标准核苷酸的三磷酸接触来扩增寡核苷酸的过程,其形成适合标准Watson-Crick几何形状的核碱基对,但是通过与加入标准A: T和G:C对。 因此,本发明涉及核苷酸类似物及其衍生物,其在掺入DNA和RNA时扩大超过标准DNA和RNA中发现的四种可重复核苷酸的数量。 本发明还涉及使用相应的三磷酸衍生物,更具体地,支持聚合酶链式反应(PCR)的聚合酶和非标准核苷三磷酸(包括产物含有的PCR)的寡核苷酸产物,将这些非标准核苷酸类似物结合到聚合酶中的聚合酶 多个非标准核苷酸单位。 提供了使用6-氨基-5-硝基-3-(1'-β-2-脱氧三呋喃糖基)-2(1H) - 吡啶酮来实施非标准“小”供体 - 供体的实施例 - 受体(pyDDA)氢键图案和2-氨基-8-(r-β-D-2'-脱氧三呋喃糖基) - 咪唑并[1,2-a] -1,3,5-三嗪-4(8H) - 执行“大”受体 - 受体 - 供体(puAAD)模式。