METHODS FOR FRACTIONATING AND PROCESSING MICROPARTICLES FROM BIOLOGICAL SAMPLES AND USING THEM FOR BIOMARKER DISCOVERY
    1.
    发明申请
    METHODS FOR FRACTIONATING AND PROCESSING MICROPARTICLES FROM BIOLOGICAL SAMPLES AND USING THEM FOR BIOMARKER DISCOVERY 审中-公开
    从生物样品中分离和处理微生物并将其用于生物识别发现的方法

    公开(公告)号:US20130052647A1

    公开(公告)日:2013-02-28

    申请号:US13578464

    申请日:2011-02-10

    摘要: Described herein are methods, devices, and compositions for fractionation and processing of microparticles from biological samples, and to methods for obtaining and using the microparticles for biomarker discovery. Biological samples include cell-free fluids, for example blood plasma, blood serum, cerebrospinal fluid, urine, and saliva, as well as conditioned media. Conditioned media is the liquid growth media used to propagate cells in vitro. Purification of microparticles from cell-free fluids is challenging, typically accomplished by prolonged ultracentrifugation. Described herein is an alternative method for efficiently harvesting and processing microparticles from cell-free fluids and from conditioned media. Embodiments described herein relate to use of the microparticles and their contents recovered from conditioned media derived from propagation of human and animal cells, as a source of biomarkers for diagnosis and prognosis of diseases and pathological conditions.

    摘要翻译: 本文描述了用于从生物样品中分离和加工微粒的方法,装置和组合物,以及获得和使用微粒用于生物标志物发现的方法。 生物样品包括无细胞流体,例如血浆,血清,脑脊髓液,尿液和唾液以及条件培养基。 条件培养基是用于在体外繁殖细胞的液体生长培养基。 从无细胞流体中纯化微粒是有挑战性的,通常通过长时间超速离心来实现。 本文描述的是用于从无细胞流体和条件培养基有效收获和加工微粒的替代方法。 本文所述的实施方案涉及从源自人和动物细胞繁殖的条件培养基中回收的微粒及其内容物作为疾病和病理状态的诊断和预后的生物标志物的来源。

    Methods for the recovery of nucleic acids from reaction mixtures
    4.
    发明授权
    Methods for the recovery of nucleic acids from reaction mixtures 失效
    从反应混合物中回收核酸的方法

    公开(公告)号:US5422241A

    公开(公告)日:1995-06-06

    申请号:US724423

    申请日:1991-07-03

    CPC分类号: C12N15/1003 C12Q1/6806

    摘要: A method for the recovery of nucleic acids from a reaction mixture, such as nuclease protection assays, comprising the use of one reagent containing a) nuclease for digesting single-stranded RNA and b) a nucleic acid precipitating carrier agent, (sheared DNA or linear acrylamide). A second reagent contains a chaotropic agent (a guanidinium salt) for inactivating said nucleases and an alcohol (ethanol or isopropanol) for the simultaneous inactivation of the nucleases and the precipitation of the nucleic acids without the need for protease digestion or organic extraction.

    摘要翻译: 用于从反应混合物中回收核酸的方法,例如核酸酶保护测定法,其包括使用含有a)核酸酶以消化单链RNA的一种试剂和b)核酸沉淀载体试剂(剪切DNA或线性 丙烯酰胺)。 第二试剂包含用于灭活所述核酸酶的离液剂(胍盐)和用于同时灭活核酸酶和沉淀核酸而不需要蛋白酶消化或有机提取的醇(乙醇或异丙醇)。

    Compositions and methods for increasing the yields of in vitro RNA
transcription and other polynucleotide synthetic reactions
    5.
    发明授权
    Compositions and methods for increasing the yields of in vitro RNA transcription and other polynucleotide synthetic reactions 失效
    用于增加体外RNA转录和其他多核苷酸合成反应产率的组合物和方法

    公开(公告)号:US5256555A

    公开(公告)日:1993-10-26

    申请号:US810968

    申请日:1991-12-20

    IPC分类号: C12P19/34 C12N9/14 C07H15/12

    CPC分类号: C12P19/34

    摘要: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA transcription and in various other enzymatic reactions in which a polynucleotide is synthesized. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

    摘要翻译: 本发明涉及增加多核苷酸合成反应产率的组合物和方法。 特别地,本发明涉及用于体外RNA转录的改进的反应混合物和其中合成多核苷酸的各种其它酶反应。 反应混合物使用高浓度的总核苷酸,大约12mM至40mM,即先前认为是抑制性的水平。 还公开了其它有用的修饰。

    METHODS AND COMPOSITIONS FOR IMPROVING REMOVAL OF RIBOSOMAL RNA FROM BIOLOGICAL SAMPLES
    7.
    发明申请
    METHODS AND COMPOSITIONS FOR IMPROVING REMOVAL OF RIBOSOMAL RNA FROM BIOLOGICAL SAMPLES 审中-公开
    改进从生物样品中除去RIBOSOMAL RNA的方法和组合物

    公开(公告)号:US20140295418A1

    公开(公告)日:2014-10-02

    申请号:US14040117

    申请日:2013-09-27

    IPC分类号: C12Q1/68

    CPC分类号: C12Q1/6806 C12Q2539/101

    摘要: The invention generally relates to compositions for maximizing capture of affinity-labeled molecules on solid supports. The disclosed methods and compositions were developed to maximize depletion of ribosomal RNA from total RNA samples, which is useful to improve the quality of RNA preparations used for applications such as massively parallel sequencing. The RNA depletion method is based on using long affinity-labeled RNA molecules that are complementary to all or part of the target ribosomal RNAs, as subtractive hybridization probes.

    摘要翻译: 本发明一般涉及使固体支持物上的亲和标记分子的捕获最大化的组合物。 开发了所公开的方法和组合物以使来自总RNA样品的核糖体RNA的消耗最大化,其可用于改善用于诸如大规模并行测序的应用的RNA制剂的质量。 RNA耗尽方法基于使用与全部或部分目标核糖体RNA互补的长亲和力标记的RNA分子作为消减杂交探针。

    OLIGONUCLEOTIDE LIGATION, BARCODING AND METHODS AND COMPOSITIONS FOR IMPROVING DATA QUALITY AND THROUGHPUT USING MASSIVELY PARALLEL SEQUENCING
    8.
    发明申请
    OLIGONUCLEOTIDE LIGATION, BARCODING AND METHODS AND COMPOSITIONS FOR IMPROVING DATA QUALITY AND THROUGHPUT USING MASSIVELY PARALLEL SEQUENCING 有权
    用于改善数据质量和使用大规模并行序列的方法和方法及组合物

    公开(公告)号:US20120028814A1

    公开(公告)日:2012-02-02

    申请号:US13102797

    申请日:2011-05-06

    IPC分类号: C40B20/00 C40B50/06

    摘要: Described herein is a buffer concentration for highly efficient ligation of two oligonucleotides. The embodiments herein have led to the development of an optimized ligation step used in the sample preparation for sequencing reactions. Further, embodiments herein describe a high-throughput method for sequencing using barcodes or the purpose of multiplexing several samples simultaneously and novel methods for making targeted DNA libraries for re-sequencing on massively parallel next-generation sequencing platforms and for alternatives to gel-purification for recovering the desired templates from small RNA libraries for next generation sequencing.

    摘要翻译: 本文描述了两种寡核苷酸的高效连接的缓冲液浓度。 本文的实施例已经导致开发用于测序反应的样品制备中的优化的连接步骤。 此外,本文的实施方案描述了使用条形码进行测序的高通量方法或同时复用多个样品的目的,以及用于制备用于大规模并行的下一代测序平台上重新测序的靶向DNA文库的新方法以及凝胶纯化替代方案 从小RNA文库中恢复所需的模板用于下一代测序。