公开(公告)号：US20170175182A1

公开(公告)日：2017-06-22

申请号：US15290622

申请日：2016-10-11

Applicant: AGILENT TECHNOLOGIES, INC.

Inventor： Brian Jon Peter ,  Robert A. Ach ,  Alicia Scheffer-Wong

IPC: C12Q1/68

CPC classification number: C12Q1/6874 ,  C12Q1/6869 ,  C12Q2521/507 ,  C12Q2565/514

Abstract: Provided herein, among other things, are a variety of methods that comprise inserting a plurality of barcoded transposons into a population of DNA fragments that comprise DNA fragments of less than 1 kb in length, to produce transposon-tagged fragments that each comprise a barcoded transposon. Kits for performing this method are also provided.

公开(公告)号：US20150125862A1

公开(公告)日：2015-05-07

申请号：US14590869

申请日：2015-01-06

Applicant: Agilent Technologies, Inc.

Inventor： Brian Phillip Smart ,  Robert A. Ach

IPC: C12Q1/68

CPC classification number: C12Q1/6876 ,  C12Q1/6844 ,  C12Q2600/158 ,  C12Q2600/178 ,  G01N33/5308 ,  G01N33/6803 ,  G01N33/6848 ,  Y10T436/143333

Abstract: Provided herein is a method of sample analysis. In certain embodiments, the method comprises: a) cross-linking protein of a cell using a first compound to produce a first cross-linked product comprising cross-linked protein, and RNA; b) contacting the first cross-linked product and a second compound under conditions by which an oligonucleotide portion of the second compound hybridizes to the RNA; c) activating a reaction the first and second compound, thereby covalently crosslinking the oligonucleotide to the cross-linked protein to produce a second cross-linked product; d) isolating the second cross-linked product using an affinity tag; and e) analyzing the isolated second cross-linked product. Compounds for performing the method are also provided.

Abstract translation: 本文提供了一种样品分析方法。 在某些实施方案中，所述方法包括：a）使用第一化合物交联细胞的蛋白质以产生包含交联蛋白质和RNA的第一交联产物; b）在第二化合物的寡核苷酸部分与RNA杂交的条件下使第一交联产物和第二化合物接触; c）使第一和第二化合物活化反应，由此共价交联寡核苷酸与交联蛋白质以产生第二交联产物; d）使用亲和标签分离第二交联产物; 和e）分析分离的第二交联产物。 还提供了用于执行该方法的化合物。

公开(公告)号：US20140357523A1

公开(公告)日：2014-12-04

申请号：US14290896

申请日：2014-05-29

Applicant: Agilent Technologies, Inc.

Inventor： Gusti Zeiner ,  Derek Lee Lindstrom ,  Brian Jon Peter ,  Robert A. Ach

IPC: C12Q1/68

CPC classification number: C12Q1/6806 ,  C12N9/22 ,  C12N15/1003 ,  C12N15/1034 ,  C12Q1/6869 ,  C12Y301/00 ,  Y10T436/143333

Abstract: A method for fragmenting a genome is provided. In certain embodiments, the method comprises: (a) combining a genomic sample containing genomic DNA with a plurality of Cas9-gRNA complexes, wherein the Cas9-gRNA complexes comprise a Cas9 protein and a set of at least 10 Cas9-associated guide RNAs that are complementary to different, pre-defined, sites in a genome, to produce a reaction mixture; and (b) incubating the reaction mixture to produce at least 5 fragments of the genomic DNA. Also provided is a composition comprising at least 100 Cas9-associated guide RNAs that are each complementary to a different, pre-defined, site in a genome. Kits for performing the method are also provided. In addition, other methods, compositions and kits for manipulating nucleic acids are also provided.

Abstract translation: 提供了一种分割基因组的方法。 在某些实施方案中，所述方法包括：（a）将含有基因组DNA的基因组样品与多个Cas9-gRNA复合物组合，其中所述Cas9-gRNA复合物包含Cas9蛋白和至少10个Cas9相关导向RNA的集合，其中 与基因组中不同的，预定义的位点互补，以产生反应混合物; 和（b）孵育反应混合物以产生至少5个基因组DNA片段。 还提供了包含至少100种与基因组中不同的，预定义的位点互补的Cas9相关导向RNA的组合物。 还提供了用于执行该方法的套件。 此外，还提供用于操作核酸的其它方法，组合物和试剂盒。

公开(公告)号：US08815782B2

公开(公告)日：2014-08-26

申请号：US13666752

申请日：2012-11-01

Applicant: Agilent Technologies, Inc.

Inventor： Gusti Zeiner ,  Robert A. Ach

IPC: C40B50/06

CPC classification number: C12P19/34 ,  C12N15/111 ,  C12N15/113 ,  C12N2310/127 ,  C12N2320/10 ,  C12Q1/6806 ,  C12Q2521/107 ,  C12Q2521/119 ,  C12Q2521/345 ,  C12Q2521/501 ,  C12Q2565/501

Abstract: Provided herein is method comprising: contacting an initial RNA sample containing a population of different RNA molecules with a divalent cation and a set of DNAzymes that are designed to cleave multiple target RNAs in the initial sample, thereby producing a product RNA sample that comprises: a) uncleaved RNA molecules and b) cleaved RNA fragments that contain a 2′,3′-cyclic-phosphate and a 5′ hydroxyl as the result of DNAzyme cleavage.

Abstract translation: 本文提供的方法包括：将含有不同RNA分子群的初始RNA样品与二价阳离子和一组设计用于在初始样品中切割多个靶RNA的DNA酶组合，从而产生产物RNA样品，其包含： ）未切割的RNA分子，以及b）由于DNA酶切割而含有2'，3'-环磷酸酯和5'羟基的切割的RNA片段。