JP6704563B2 - カオトロープ剤及びカオトロープ剤を用いてゲノムｄｎａを抽出する方法 - Google Patents
カオトロープ剤及びカオトロープ剤を用いてゲノムｄｎａを抽出する方法 Download PDFInfo
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- JP6704563B2 JP6704563B2 JP2019515850A JP2019515850A JP6704563B2 JP 6704563 B2 JP6704563 B2 JP 6704563B2 JP 2019515850 A JP2019515850 A JP 2019515850A JP 2019515850 A JP2019515850 A JP 2019515850A JP 6704563 B2 JP6704563 B2 JP 6704563B2
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- genomic dna
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- test tube
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- 238000000034 method Methods 0.000 title claims description 80
- XUXNAKZDHHEHPC-UHFFFAOYSA-M sodium bromate Chemical group [Na+].[O-]Br(=O)=O XUXNAKZDHHEHPC-UHFFFAOYSA-M 0.000 claims description 39
- 108010033040 Histones Proteins 0.000 claims description 31
- 239000003795 chemical substances by application Substances 0.000 claims description 26
- 230000003196 chaotropic effect Effects 0.000 claims description 25
- 239000006228 supernatant Substances 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- 239000000203 mixture Substances 0.000 claims description 20
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 18
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 14
- 238000012360 testing method Methods 0.000 claims description 13
- 239000013592 cell lysate Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 10
- 239000002244 precipitate Substances 0.000 claims description 9
- 210000004369 blood Anatomy 0.000 claims description 7
- 239000008280 blood Substances 0.000 claims description 7
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 5
- 239000007984 Tris EDTA buffer Substances 0.000 claims description 5
- SXDBWCPKPHAZSM-UHFFFAOYSA-M bromate Inorganic materials [O-]Br(=O)=O SXDBWCPKPHAZSM-UHFFFAOYSA-M 0.000 claims description 4
- SXDBWCPKPHAZSM-UHFFFAOYSA-N bromic acid Chemical compound OBr(=O)=O SXDBWCPKPHAZSM-UHFFFAOYSA-N 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 108010067770 Endopeptidase K Proteins 0.000 description 39
- 210000002919 epithelial cell Anatomy 0.000 description 18
- 230000029087 digestion Effects 0.000 description 14
- CRWJEUDFKNYSBX-UHFFFAOYSA-N sodium;hypobromite Chemical compound [Na+].Br[O-] CRWJEUDFKNYSBX-UHFFFAOYSA-N 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 12
- 210000003296 saliva Anatomy 0.000 description 12
- BAZAXWOYCMUHIX-UHFFFAOYSA-M sodium perchlorate Chemical compound [Na+].[O-]Cl(=O)(=O)=O BAZAXWOYCMUHIX-UHFFFAOYSA-M 0.000 description 12
- 229910001488 sodium perchlorate Inorganic materials 0.000 description 12
- 102000006947 Histones Human genes 0.000 description 11
- 238000000605 extraction Methods 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- 238000001514 detection method Methods 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 238000000246 agarose gel electrophoresis Methods 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 238000000424 optical density measurement Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 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 3
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 3
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 3
- 238000004925 denaturation Methods 0.000 description 3
- 230000036425 denaturation Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- MHCFAGZWMAWTNR-UHFFFAOYSA-M lithium perchlorate Chemical compound [Li+].[O-]Cl(=O)(=O)=O MHCFAGZWMAWTNR-UHFFFAOYSA-M 0.000 description 3
- 229910001486 lithium perchlorate Inorganic materials 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 210000002200 mouth mucosa Anatomy 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 241000206602 Eukaryota Species 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 2
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-M perchlorate Inorganic materials [O-]Cl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-M 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 230000004544 DNA amplification Effects 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
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
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710413739.9 | 2017-06-05 | ||
| CN201710413739.9A CN107058297B (zh) | 2017-06-05 | 2017-06-05 | 一种离液剂以及使用该离液剂提取基因组dna的方法 |
| PCT/CN2017/116735 WO2018223657A1 (zh) | 2017-06-05 | 2017-12-17 | 一种离液剂以及使用该离液剂提取基因组dna的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2019528746A JP2019528746A (ja) | 2019-10-17 |
| JP6704563B2 true JP6704563B2 (ja) | 2020-06-03 |
Family
ID=59616361
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2019515850A Active JP6704563B2 (ja) | 2017-06-05 | 2017-12-17 | カオトロープ剤及びカオトロープ剤を用いてゲノムｄｎａを抽出する方法 |
Country Status (3)
| Country | Link |
|---|---|
| JP (1) | JP6704563B2 (zh) |
| CN (1) | CN107058297B (zh) |
| WO (1) | WO2018223657A1 (zh) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058297B (zh) * | 2017-06-05 | 2018-01-16 | 陈礼平 | 一种离液剂以及使用该离液剂提取基因组dna的方法 |
| CN109694863B (zh) * | 2019-02-28 | 2021-04-20 | 深圳市刚竹医疗科技有限公司 | 用于核酸提取的裂解液、清洗液、核酸提取试剂盒及核酸的提取方法 |
| CN113308460A (zh) * | 2020-12-15 | 2021-08-27 | 深圳未知君生物科技有限公司 | 一种提取石蜡切片中细菌dna的试剂盒 |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5733462A (en) * | 1996-10-03 | 1998-03-31 | Cytec Technology Corp. | Cationic water-soluble polymer precipitation in salt solutions |
| US5696228A (en) * | 1996-10-03 | 1997-12-09 | Cytec Technology Corp. | Process for producing substantially dry polymer particles from aqueous dispersions |
| US7794932B2 (en) * | 2004-04-16 | 2010-09-14 | Piotr Chomczynski | Reagents and methods for isolation of purified RNA |
| US7867592B2 (en) * | 2007-01-30 | 2011-01-11 | Eksigent Technologies, Inc. | Methods, compositions and devices, including electroosmotic pumps, comprising coated porous surfaces |
| EP2729570B1 (en) * | 2011-07-04 | 2017-11-22 | Qiagen GmbH | Reagent usable for the isolation and/or purification of nucleic acids |
| CN107058297B (zh) * | 2017-06-05 | 2018-01-16 | 陈礼平 | 一种离液剂以及使用该离液剂提取基因组dna的方法 |
-
2017
- 2017-06-05 CN CN201710413739.9A patent/CN107058297B/zh active Active
- 2017-12-17 WO PCT/CN2017/116735 patent/WO2018223657A1/zh active Application Filing
- 2017-12-17 JP JP2019515850A patent/JP6704563B2/ja active Active
Also Published As
| Publication number | Publication date |
|---|---|
| JP2019528746A (ja) | 2019-10-17 |
| CN107058297A (zh) | 2017-08-18 |
| WO2018223657A1 (zh) | 2018-12-13 |
| CN107058297B (zh) | 2018-01-16 |
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