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1. WO2020191521 - NUCLEOTIDE SEQUENCE, AND METHOD FOR CONSTRUCTING RNA TARGET AREA SEQUENCING LIBRARY AND APPLICATION THEREOF

Publication Number WO/2020/191521
Publication Date 01.10.2020
International Application No. PCT/CN2019/079235
International Filing Date 22.03.2019
IPC
C12N 15/11 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C40B 50/06 2006.01
CCHEMISTRY; METALLURGY
40COMBINATORIAL TECHNOLOGY
BCOMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICO LIBRARIES
50Methods of creating libraries, e.g. combinatorial synthesis
06Biochemical methods, e.g. using enzymes or whole viable microorganisms
C12Q 1/6806 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction assay
C12Q 1/6809 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6809Methods for determination or identification of nucleic acids involving differential detection
C12Q 1/6886 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
6883for diseases caused by alterations of genetic material
6886for cancer
CPC
C12N 15/11
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C12Q 1/6806
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6809
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6809Methods for determination or identification of nucleic acids involving differential detection
C12Q 1/6886
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6876Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
6883for diseases caused by alterations of genetic material
6886for cancer
C40B 50/06
CCHEMISTRY; METALLURGY
40COMBINATORIAL TECHNOLOGY
BCOMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICO LIBRARIES
50Methods of creating libraries, e.g. combinatorial synthesis
06Biochemical methods, e.g. using enzymes or whole viable microorganisms
Applicants
  • 深圳华大智造科技有限公司 MGI TECH CO., LTD. [CN]/[CN]
Inventors
  • 杨林 YANG, Lin
  • 夏军 XIA, Jun
  • 张艳艳 ZHANG, Yanyan
  • 吕硕 LV, Shuo
  • 陈芳 CHEN, Fang
  • 梅智颖 MEI, Zhiying
  • 陈大洋 CHEN, Dayang
  • 蒋慧 JIANG, Hui
Agents
  • 北京清亦华知识产权代理事务所(普通合伙) TSINGYIHUA INTELLECTUAL PROPERTY LLC
Priority Data
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) NUCLEOTIDE SEQUENCE, AND METHOD FOR CONSTRUCTING RNA TARGET AREA SEQUENCING LIBRARY AND APPLICATION THEREOF
(FR) SÉQUENCE NUCLÉOTIDIQUE, ET PROCÉDÉ DE CONSTRUCTION D'UNE BANQUE DE SÉQUENÇAGE DE ZONE CIBLE D'ARN ET APPLICATION CORRESPONDANTE
(ZH) 核酸序列、RNA目标区域测序文库的构建方法及应用
Abstract
(EN)
Provided are a nucleotide sequence, and a method for constructing an RNA target area sequencing library and an application thereof. The nucleotide sequence comprises a first general sequencing sequence located at terminus 5' and a random primer sequence located at terminus 3' which are connected to each other. The random primer sequence contains 6-10 random nucleotides. The sequence is used to perform random reverse transcription on RNA to obtain a first chain of cDNA, and then the first general sequencing sequence on the nucleotide sequence and upstream and downstream specific primers are used to perform two-step nested PCR on the first chain of cDNA obtained after the reverse transcription, to obtain a high-throughput targeted sequencing library.
(FR)
L'invention concerne une séquence nucléotidique, et un procédé de construction d'une bibliothèque de séquençage de zone cible d'ARN et une application correspondante. La séquence nucléotidique comprend une première séquence de séquençage générale située à l'extrémité 5' et une séquence d'amorce aléatoire située au niveau de l'extrémité 3' étant reliées l'une à l'autre. La séquence d'amorce aléatoire contient de 6 à 10 nucléotides aléatoires. La séquence est utilisée pour effectuer une transcription inverse aléatoire sur l'ARN pour obtenir une première chaîne d'ADNc, puis la première séquence de séquençage générale sur la séquence de nucléotides et des amorces spécifiques amont et aval sont utilisées pour effectuer une PCR nichée en deux étapes sur la première chaîne d'ADNc obtenue après la transcription inverse, pour obtenir une banque de séquençage ciblée à haut débit.
(ZH)
提供了一种核酸序列以及RNA目标区域测序文库的构建方法及应用。该核酸序列包含相连的位于5'端的第一测序通用序列和位于3'端的随机引物序列,随机引物序列含有6~10个随机核苷酸。利用该序列对RNA进行随机的逆转录,得到cDNA的第一链,然后通过该核酸序列上的第一测序通用序列以及上下游特异性引物对逆转录得到的cDNA第一条链进行2步巢式PCR,得到高通量靶向测序文库。
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