CN112601825A - 核酸测序方法 - Google Patents
核酸测序方法 Download PDFInfo
- Publication number
- CN112601825A CN112601825A CN201880094670.0A CN201880094670A CN112601825A CN 112601825 A CN112601825 A CN 112601825A CN 201880094670 A CN201880094670 A CN 201880094670A CN 112601825 A CN112601825 A CN 112601825A
- Authority
- CN
- China
- Prior art keywords
- nucleotide
- group
- nucleic acid
- signal
- nucleotide analog
- 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
- 238000003203 nucleic acid sequencing method Methods 0.000 title abstract description 6
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 275
- 238000012163 sequencing technique Methods 0.000 claims abstract description 110
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 85
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 72
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 72
- 238000000034 method Methods 0.000 claims abstract description 71
- 239000002773 nucleotide Substances 0.000 claims abstract description 45
- 238000006243 chemical reaction Methods 0.000 claims description 87
- 239000000203 mixture Substances 0.000 claims description 45
- 230000003287 optical effect Effects 0.000 claims description 40
- 230000005284 excitation Effects 0.000 claims description 34
- 230000000903 blocking effect Effects 0.000 claims description 29
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims description 24
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 24
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims description 24
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 24
- 108020004414 DNA Proteins 0.000 claims description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 claims description 14
- 229930024421 Adenine Natural products 0.000 claims description 12
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 12
- 229960000643 adenine Drugs 0.000 claims description 12
- 229940104302 cytosine Drugs 0.000 claims description 12
- 229940113082 thymine Drugs 0.000 claims description 12
- 229940035893 uracil Drugs 0.000 claims description 12
- 238000003776 cleavage reaction Methods 0.000 claims description 10
- 230000007017 scission Effects 0.000 claims description 10
- 125000006239 protecting group Chemical group 0.000 claims description 9
- ASJSAQIRZKANQN-CRCLSJGQSA-N 2-deoxy-D-ribose Chemical group OC[C@@H](O)[C@@H](O)CC=O ASJSAQIRZKANQN-CRCLSJGQSA-N 0.000 claims description 8
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 claims description 8
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 claims description 8
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 7
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 6
- 230000011664 signaling Effects 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 5
- 239000012071 phase Substances 0.000 claims description 5
- 230000003197 catalytic effect Effects 0.000 claims description 4
- 230000000295 complement effect Effects 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000000137 annealing Methods 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 239000000376 reactant Substances 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- 238000004925 denaturation Methods 0.000 claims description 2
- 230000036425 denaturation Effects 0.000 claims description 2
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 claims 1
- 239000013615 primer Substances 0.000 description 19
- 150000001875 compounds Chemical class 0.000 description 18
- 238000001712 DNA sequencing Methods 0.000 description 14
- 239000007850 fluorescent dye Substances 0.000 description 14
- 230000008569 process Effects 0.000 description 12
- 238000005516 engineering process Methods 0.000 description 11
- 238000003559 RNA-seq method Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000003752 polymerase chain reaction Methods 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 238000004020 luminiscence type Methods 0.000 description 4
- 230000003321 amplification Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 238000007480 sanger sequencing Methods 0.000 description 3
- 125000000171 (C1-C6) haloalkyl group Chemical group 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 229920000388 Polyphosphate Chemical group 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- XPPKVPWEQAFLFU-UHFFFAOYSA-N diphosphoric acid Chemical group OP(O)(=O)OP(O)(O)=O XPPKVPWEQAFLFU-UHFFFAOYSA-N 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 150000004712 monophosphates Chemical group 0.000 description 2
- 239000001205 polyphosphate Chemical group 0.000 description 2
- 235000011176 polyphosphates Nutrition 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 238000007841 sequencing by ligation Methods 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 125000002264 triphosphate group Chemical group [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 108020001019 DNA Primers Proteins 0.000 description 1
- 238000000018 DNA microarray Methods 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Natural products P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 1
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical class O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000012177 large-scale sequencing Methods 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- -1 phosphine compound Chemical class 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000006916 protein interaction Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Images
Classifications
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Wood Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明提供了一种核酸测序方法以及相应的测序装置，所述方法特征在于扩增待测核酸分子双链体时将带有荧光和磷光基团的核苷酸类似物并入起始生长核酸链的3'端，通过检测荧光和磷光信号，判断待测核酸分子3'端的核苷酸。
Description
PCT国内申请，说明书已公开。
Claims (26)
- PCT国内申请，权利要求书已公开。
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2018/095040 WO2020010495A1 (zh) | 2018-07-09 | 2018-07-09 | 核酸测序方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112601825A true CN112601825A (zh) | 2021-04-02 |
| CN112601825B CN112601825B (zh) | 2023-11-28 |
Family
ID=69142276
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201880094670.0A Active CN112601825B (zh) | 2018-07-09 | 2018-07-09 | 核酸测序方法 |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN112601825B (zh) |
| WO (1) | WO2020010495A1 (zh) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114250283A (zh) * | 2021-10-15 | 2022-03-29 | 深圳铭毅智造科技有限公司 | 基于环境敏感染料的单色荧光mrt基因测序试剂及方法 |
| CN114250281A (zh) * | 2021-11-03 | 2022-03-29 | 深圳铭毅智造科技有限公司 | 一种核酸代谢酶活性检测方法 |
| CN114958995A (zh) * | 2022-04-27 | 2022-08-30 | 深圳赛陆医疗科技有限公司 | 一种基因测序方法 |
| CN115232867A (zh) * | 2022-07-21 | 2022-10-25 | 深圳赛陆医疗科技有限公司 | 一种快速的二代基因测序方法 |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN119061122A (zh) * | 2023-05-30 | 2024-12-03 | 深圳市真迈生物科技有限公司 | 单分子测序方法 |
| WO2025007252A1 (zh) * | 2023-07-03 | 2025-01-09 | 深圳华大智造科技股份有限公司 | 重复测序的方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070117104A1 (en) * | 2005-11-22 | 2007-05-24 | Buzby Philip R | Nucleotide analogs |
| JP2012090546A (ja) * | 2010-10-26 | 2012-05-17 | Shimadzu Corp | ラベル化ヌクレオチドの精製方法及びそれを用いる核酸配列解読装置 |
| WO2018060482A1 (en) * | 2016-09-30 | 2018-04-05 | Illumina Cambridge Limited | New fluorescent dyes and their uses as biomarkers |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6031091A (en) * | 1987-09-21 | 2000-02-29 | Gen-Probe Incorporated | Non-nucleotide linking reagents for nucleotide probes |
| WO2003052133A2 (en) * | 2001-12-18 | 2003-06-26 | Human Genetic Signatures Pty Ltd | Oligonucleotide analogues comprising intercalator pseudonucleotides |
| AU2003277984A1 (en) * | 2002-11-05 | 2004-06-07 | The University Of Queensland | Nucleotide sequence analysis by quantification of mutagenesis |
| GB2398383B (en) * | 2003-02-12 | 2005-03-09 | Global Genomics Ab | Method and means for nucleic acid sequencing |
| WO2006063717A2 (en) * | 2004-12-16 | 2006-06-22 | Febit Biotech Gmbh | Polymerase-independent analysis of the sequence of polynucleotides |
-
2018
- 2018-07-09 CN CN201880094670.0A patent/CN112601825B/zh active Active
- 2018-07-09 WO PCT/CN2018/095040 patent/WO2020010495A1/zh not_active Ceased
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070117104A1 (en) * | 2005-11-22 | 2007-05-24 | Buzby Philip R | Nucleotide analogs |
| JP2012090546A (ja) * | 2010-10-26 | 2012-05-17 | Shimadzu Corp | ラベル化ヌクレオチドの精製方法及びそれを用いる核酸配列解読装置 |
| WO2018060482A1 (en) * | 2016-09-30 | 2018-04-05 | Illumina Cambridge Limited | New fluorescent dyes and their uses as biomarkers |
Non-Patent Citations (1)
| Title |
|---|
| G. ZHANG等: "A dual-emissive-materials design concept enables tumour hypoxia imaging", NATURE MATER, vol. 8, no. 9, pages 1 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114250283A (zh) * | 2021-10-15 | 2022-03-29 | 深圳铭毅智造科技有限公司 | 基于环境敏感染料的单色荧光mrt基因测序试剂及方法 |
| CN114250281A (zh) * | 2021-11-03 | 2022-03-29 | 深圳铭毅智造科技有限公司 | 一种核酸代谢酶活性检测方法 |
| CN114958995A (zh) * | 2022-04-27 | 2022-08-30 | 深圳赛陆医疗科技有限公司 | 一种基因测序方法 |
| WO2023207265A1 (zh) * | 2022-04-27 | 2023-11-02 | 深圳赛陆医疗科技有限公司 | 一种基因测序方法 |
| CN115232867A (zh) * | 2022-07-21 | 2022-10-25 | 深圳赛陆医疗科技有限公司 | 一种快速的二代基因测序方法 |
| CN115232867B (zh) * | 2022-07-21 | 2024-01-30 | 深圳赛陆医疗科技有限公司 | 一种快速的二代基因测序方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112601825B (zh) | 2023-11-28 |
| WO2020010495A1 (zh) | 2020-01-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112601825B (zh) | 核酸测序方法 | |
| US9938573B2 (en) | Methods and kits for nucleic acid sequencing | |
| CN106434871B (zh) | 用于检测目标核酸的方法与组合物 | |
| EP3623481B1 (en) | Compositions for nucleic acid sequencing | |
| RU2609630C2 (ru) | Геномный отбор и секвенирование с помощью кодированных микроносителей | |
| CN112840035B (zh) | 对多核苷酸进行测序的方法 | |
| KR20150060987A (ko) | 생물학적 프로브 및 이의 용도 | |
| JP6510978B2 (ja) | 核酸を配列決定する方法および装置 | |
| US7049104B2 (en) | Genetic analysis method | |
| JP2022533916A (ja) | 自家発光に基づく単一チャネルシーケンシング法 | |
| CN114286867B (zh) | 一种基于发光标记物光信号动力学及二次发光信号对多核苷酸进行测序的方法 | |
| EP1384789A1 (en) | Fluorescent hybridization probes with reduced background | |
| CN115803457A (zh) | 用于核酸测序的方法、系统和组合物 | |
| JP2025538399A (ja) | Ｐｃｒ法における相補一本鎖蛍光レポーター複合体の使用 | |
| CN117625767A (zh) | 一种文库扩增单克隆纯度表征方法 | |
| HK40049621A (zh) | 对多核苷酸进行测序的方法 | |
| WO2011108344A1 (ja) | 基板上に固定化された複数の核酸検体の識別方法及び装置 | |
| HK1198548B (zh) | 用於检测目标核酸的方法与组合物 |
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 | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20231030 Address after: Building 4, No. 2 Hengyunshan Road, Qingdao Area, China (Shandong) Pilot Free Trade Zone, Qingdao City, Shandong Province, 266426 Applicant after: Qingdao Huada Zhizao Technology Co.,Ltd. Address before: 518083 7th floor, building 11, Beishan Industrial Zone, 146 Beishan Road, Yangang community, Yantian street, Yantian District, Shenzhen City, Guangdong Province Applicant before: Shenzhen Huada zhizaojichuang Technology Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |