CN113227370A - 一种单链dna合成方法 - Google Patents
一种单链dna合成方法 Download PDFInfo
- Publication number
- CN113227370A CN113227370A CN201980086128.5A CN201980086128A CN113227370A CN 113227370 A CN113227370 A CN 113227370A CN 201980086128 A CN201980086128 A CN 201980086128A CN 113227370 A CN113227370 A CN 113227370A
- Authority
- CN
- China
- Prior art keywords
- sequence
- dna
- stranded dna
- stranded
- uracil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108020004414 DNA Proteins 0.000 title claims abstract description 97
- 102000053602 DNA Human genes 0.000 title claims abstract description 79
- 108020004682 Single-Stranded DNA Proteins 0.000 title claims abstract description 58
- 230000006820 DNA synthesis Effects 0.000 title abstract description 3
- 238000001308 synthesis method Methods 0.000 title abstract description 3
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims abstract description 46
- 102000004190 Enzymes Human genes 0.000 claims abstract description 26
- 108090000790 Enzymes Proteins 0.000 claims abstract description 26
- 230000010076 replication Effects 0.000 claims abstract description 25
- 238000005096 rolling process Methods 0.000 claims abstract description 23
- 229940035893 uracil Drugs 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims description 53
- 108091008146 restriction endonucleases Proteins 0.000 claims description 52
- 238000006243 chemical reaction Methods 0.000 claims description 36
- 239000002773 nucleotide Substances 0.000 claims description 27
- 125000003729 nucleotide group Chemical group 0.000 claims description 27
- 238000003776 cleavage reaction Methods 0.000 claims description 21
- 230000007017 scission Effects 0.000 claims description 21
- 230000000295 complement effect Effects 0.000 claims description 19
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 18
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 18
- 239000003153 chemical reaction reagent Substances 0.000 claims description 16
- 102000012410 DNA Ligases Human genes 0.000 claims description 15
- 108010061982 DNA Ligases Proteins 0.000 claims description 15
- 102000003960 Ligases Human genes 0.000 claims description 12
- 108090000364 Ligases Proteins 0.000 claims description 12
- 238000005520 cutting process Methods 0.000 claims description 12
- 238000012408 PCR amplification Methods 0.000 claims description 11
- 238000000137 annealing Methods 0.000 claims description 11
- 108091023037 Aptamer Proteins 0.000 claims description 10
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 9
- 230000003321 amplification Effects 0.000 claims description 7
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 7
- 238000012300 Sequence Analysis Methods 0.000 claims description 6
- 230000006978 adaptation Effects 0.000 claims description 4
- GFFGJBXGBJISGV-UHFFFAOYSA-N adenyl group Chemical group N1=CN=C2N=CNC2=C1N GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 4
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical group CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 4
- 108010064978 Type II Site-Specific Deoxyribonucleases Proteins 0.000 claims description 2
- 238000003752 polymerase chain reaction Methods 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 101100481002 Mus musculus Tff1 gene Proteins 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 9
- 238000007363 ring formation reaction Methods 0.000 abstract description 6
- 230000001404 mediated effect Effects 0.000 abstract description 2
- 230000035772 mutation Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 48
- 229940088598 enzyme Drugs 0.000 description 23
- 239000012634 fragment Substances 0.000 description 21
- 238000000746 purification Methods 0.000 description 18
- 239000012154 double-distilled water Substances 0.000 description 14
- 239000000872 buffer Substances 0.000 description 13
- 238000000246 agarose gel electrophoresis Methods 0.000 description 12
- 239000011324 bead Substances 0.000 description 11
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 238000001816 cooling Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000178 monomer Substances 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000012165 high-throughput sequencing Methods 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 108020004638 Circular DNA Proteins 0.000 description 4
- 230000000415 inactivating effect Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 108050009160 DNA polymerase 1 Proteins 0.000 description 3
- 230000003044 adaptive effect Effects 0.000 description 3
- 239000007795 chemical reaction product Substances 0.000 description 3
- 239000012043 crude product Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 108060002716 Exonuclease Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 238000007846 asymmetric PCR Methods 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 102000013165 exonuclease Human genes 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 238000003766 bioinformatics method Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000003198 gene knock in Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000011901 isothermal amplification Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000007169 ligase reaction Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 238000012772 sequence design Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
- C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q2521/00—Reaction characterised by the enzymatic activity
- C12Q2521/30—Phosphoric diester hydrolysing, i.e. nuclease
- C12Q2521/301—Endonuclease
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q2521/00—Reaction characterised by the enzymatic activity
- C12Q2521/50—Other enzymatic activities
- C12Q2521/501—Ligase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Plant Pathology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Crystallography & Structural Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
提供了一种单链DNA合成方法，具体为一种无碱基突变的单链DNA的制备工艺，通过尿嘧啶特异性减除酶(USER)介导的双链DNA自动成环结合滚环复制来制备单链DNA。
Description
PCT国内申请，说明书已公开。
Claims (19)
- PCT国内申请，权利要求书已公开。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811624165 | 2018-12-28 | ||
| CN2018116241650 | 2018-12-28 | ||
| PCT/CN2019/128948 WO2020135651A1 (zh) | 2018-12-28 | 2019-12-27 | 一种单链dna合成方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113227370A true CN113227370A (zh) | 2021-08-06 |
| CN113227370B CN113227370B (zh) | 2024-04-09 |
Family
ID=71126836
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201980086128.5A Active CN113227370B (zh) | 2018-12-28 | 2019-12-27 | 一种单链dna合成方法 |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US20220042059A1 (zh) |
| EP (1) | EP3904511A4 (zh) |
| JP (1) | JP2022515085A (zh) |
| KR (1) | KR20210110790A (zh) |
| CN (1) | CN113227370B (zh) |
| IL (1) | IL283989A (zh) |
| TW (1) | TW202039838A (zh) |
| WO (1) | WO2020135651A1 (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022170707A1 (zh) * | 2021-02-10 | 2022-08-18 | 清华大学 | 一种制备定点修饰的长链dna的方法 |
| WO2023072145A1 (zh) | 2021-10-26 | 2023-05-04 | 南京金斯瑞生物科技有限公司 | 一种纯化单链dna的方法 |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007135354A1 (en) * | 2006-05-19 | 2007-11-29 | Plant Bioscience Limited | Improved uracil-excision based molecular cloning |
| WO2011017760A1 (en) * | 2009-08-12 | 2011-02-17 | Commonwealth Scientific And Industrial Research Organisation | Detection and analysis of methylation in nucleic acid sequences |
| CN102296065A (zh) * | 2011-08-04 | 2011-12-28 | 盛司潼 | 用于构建测序文库的系统与方法 |
| CN102719550A (zh) * | 2012-07-10 | 2012-10-10 | 中国人民解放军第三军医大学第一附属医院 | 一种基于分裂锁式探针的多重rca方法 |
| CN103255227A (zh) * | 2013-05-30 | 2013-08-21 | 上海快灵生物科技有限公司 | 引物介导环化的恒温核酸滚环扩增方法及试剂盒 |
| US20130261027A1 (en) * | 2008-01-09 | 2013-10-03 | Life Technologies Corporation | Nucleic acid adaptors and uses thereof |
| US20180044668A1 (en) * | 2014-10-14 | 2018-02-15 | Bgi Shenzhen Co., Limited | Mate pair library construction |
| CN108060191A (zh) * | 2017-11-07 | 2018-05-22 | 深圳华大基因科技有限公司 | 一种双链核酸片段加接头的方法、文库构建方法和试剂盒 |
| WO2018090373A1 (zh) * | 2016-11-21 | 2018-05-24 | 深圳华大智造科技有限公司 | 一种dna末端修复与加a的方法 |
| WO2018121634A1 (zh) * | 2016-12-30 | 2018-07-05 | 安诺优达基因科技(北京)有限公司 | 用于dna片段的非特异性复制的方法及试剂盒 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009120374A2 (en) * | 2008-03-28 | 2009-10-01 | Pacific Biosciences Of California, Inc. | Methods and compositions for nucleic acid sample preparation |
| CN105624165B (zh) * | 2016-01-05 | 2018-11-30 | 山东大学 | 基于自锁式适体探针的级联扩增策略的生物分子检测方法 |
-
2019
- 2019-12-27 TW TW108147968A patent/TW202039838A/zh unknown
- 2019-12-27 KR KR1020217014403A patent/KR20210110790A/ko unknown
- 2019-12-27 CN CN201980086128.5A patent/CN113227370B/zh active Active
- 2019-12-27 JP JP2021534996A patent/JP2022515085A/ja active Pending
- 2019-12-27 EP EP19903928.0A patent/EP3904511A4/en active Pending
- 2019-12-27 US US17/309,488 patent/US20220042059A1/en active Pending
- 2019-12-27 WO PCT/CN2019/128948 patent/WO2020135651A1/zh unknown
-
2021
- 2021-06-14 IL IL283989A patent/IL283989A/en unknown
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007135354A1 (en) * | 2006-05-19 | 2007-11-29 | Plant Bioscience Limited | Improved uracil-excision based molecular cloning |
| US20130261027A1 (en) * | 2008-01-09 | 2013-10-03 | Life Technologies Corporation | Nucleic acid adaptors and uses thereof |
| WO2011017760A1 (en) * | 2009-08-12 | 2011-02-17 | Commonwealth Scientific And Industrial Research Organisation | Detection and analysis of methylation in nucleic acid sequences |
| CN102296065A (zh) * | 2011-08-04 | 2011-12-28 | 盛司潼 | 用于构建测序文库的系统与方法 |
| CN102719550A (zh) * | 2012-07-10 | 2012-10-10 | 中国人民解放军第三军医大学第一附属医院 | 一种基于分裂锁式探针的多重rca方法 |
| CN103255227A (zh) * | 2013-05-30 | 2013-08-21 | 上海快灵生物科技有限公司 | 引物介导环化的恒温核酸滚环扩增方法及试剂盒 |
| US20180044668A1 (en) * | 2014-10-14 | 2018-02-15 | Bgi Shenzhen Co., Limited | Mate pair library construction |
| WO2018090373A1 (zh) * | 2016-11-21 | 2018-05-24 | 深圳华大智造科技有限公司 | 一种dna末端修复与加a的方法 |
| WO2018121634A1 (zh) * | 2016-12-30 | 2018-07-05 | 安诺优达基因科技(北京)有限公司 | 用于dna片段的非特异性复制的方法及试剂盒 |
| CN108060191A (zh) * | 2017-11-07 | 2018-05-22 | 深圳华大基因科技有限公司 | 一种双链核酸片段加接头的方法、文库构建方法和试剂盒 |
Non-Patent Citations (5)
| Title |
|---|
| BITINATE, J: "USER Friendly DNA Engineering and Cloning Method by Uracil Excision", NUCLEIC ACIDS RESEARCH, pages 1362 - 4962 * |
| JINFENG CHEN: "Enzyme-free synthesis of cyclic single-stranded DNA constructs containing a single triazole, amide or phosphoramidate backbone linkage and their use as templates for rolling circle amplification and nanoflower formation", CHEM. SCI, pages 8110 - 8120 * |
| M. MONSUR ALI: "Rolling circle amplification: a versatile tool for chemical biology, materials science and medicine", CHEM. SOC. REV, pages 3324 - 3341 * |
| 李启明: "滚环复制技术的建立及在RNA病毒基因检测中的初步应用", 病毒学报, 25 August 2005 (2005-08-25) * |
| 贾友礼: "基于序列信息高度集成的长单链DNA自组装及制备", 科学通报, 16 October 2018 (2018-10-16) * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2020135651A1 (zh) | 2020-07-02 |
| KR20210110790A (ko) | 2021-09-09 |
| EP3904511A1 (en) | 2021-11-03 |
| EP3904511A4 (en) | 2022-03-09 |
| US20220042059A1 (en) | 2022-02-10 |
| IL283989A (en) | 2021-07-29 |
| JP2022515085A (ja) | 2022-02-17 |
| CN113227370B (zh) | 2024-04-09 |
| TW202039838A (zh) | 2020-11-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3298170B1 (en) | Methods for generating circular dna from circular rna | |
| CN113046413A (zh) | 从血液特异性靶向捕获人类基因组和转录组区域的方法 | |
| EP2475777A1 (en) | Compositions and methods for whole transcriptome analysis | |
| JP2011500092A (ja) | 非ランダムプライマーを用いたｃＤＮＡ合成の方法 | |
| CN109593757B (zh) | 一种探针及其适用于高通量测序的对目标区域进行富集的方法 | |
| KR101600039B1 (ko) | 염기 특이 반응성 프라이머를 이용한 핵산 증폭방법 | |
| US20180195099A1 (en) | Enzymatic synthesis of nucleic acid sequences | |
| CN113227370A (zh) | 一种单链dna合成方法 | |
| CN111074353A (zh) | 全基因组甲基化文库单链建库方法和得到的全基因组甲基化文库 | |
| CN116043337A (zh) | Dna甲基化标志物筛查试剂盒及方法 | |
| WO2019185034A1 (zh) | 一种sgRNA的体外合成方法及其试剂盒 | |
| CN110699425A (zh) | 基因目标区域的富集方法及体系 | |
| CN113564197A (zh) | 一种CRISPR/Cas9介导的植物多基因编辑载体的构建方法和应用 | |
| US20210355519A1 (en) | Demand synthesis of polynucleotide sequences | |
| CN108103052B (zh) | 提高基因组覆盖度的单细胞全基因组扩增及文库构建方法 | |
| CN111893170B (zh) | 一种在全基因组范围内体外检测CRISPR-Cas脱靶效应的方法 | |
| US10450557B2 (en) | Method of nucleic acid fragmentation | |
| US20080213841A1 (en) | Novel Method for Assembling DNA Metasegments to use as Substrates for Homologous Recombination in a Cell | |
| CN116135982A (zh) | 用于基因片段连接的应用方法和试剂盒 | |
| JP7333171B2 (ja) | Ｒｎａ検出方法、ｒｎａ検出用核酸及びｒｎａ検出用キット | |
| WO2023220110A1 (en) | Highly efficient and simple ssper and rrpcr approaches for the accurate site-directed mutagenesis of large plasmids | |
| KR20240045127A (ko) | 효소 반응을 통한 총 mRNA 기반 무작위 sgRNA 라이브러리 생성 방법 | |
| CN113943779A (zh) | 一种高cg含量dna序列的富集方法及其应用 | |
| CN111455030A (zh) | 一种仅基于化学试剂的新型基因克隆方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |