HRP20201452T1 - Izoliranje nukleinskih kiselina - Google Patents
Izoliranje nukleinskih kiselina Download PDFInfo
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- HRP20201452T1 HRP20201452T1 HRP20201452TT HRP20201452T HRP20201452T1 HR P20201452 T1 HRP20201452 T1 HR P20201452T1 HR P20201452T T HRP20201452T T HR P20201452TT HR P20201452 T HRP20201452 T HR P20201452T HR P20201452 T1 HRP20201452 T1 HR P20201452T1
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- ethanol
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- 150000007523 nucleic acids Chemical class 0.000 title claims 4
- 102000039446 nucleic acids Human genes 0.000 title claims 4
- 108020004707 nucleic acids Proteins 0.000 title claims 4
- 238000002955 isolation Methods 0.000 title 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 22
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 claims 16
- 239000007788 liquid Substances 0.000 claims 13
- 239000007787 solid Substances 0.000 claims 10
- 238000000034 method Methods 0.000 claims 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 9
- 238000005406 washing Methods 0.000 claims 8
- 239000007864 aqueous solution Substances 0.000 claims 5
- 239000000243 solution Substances 0.000 claims 5
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims 4
- 159000000000 sodium salts Chemical class 0.000 claims 4
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 claims 3
- 239000012620 biological material Substances 0.000 claims 3
- 230000009089 cytolysis Effects 0.000 claims 3
- 239000000835 fiber Substances 0.000 claims 3
- 229910052744 lithium Inorganic materials 0.000 claims 3
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical class NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 claims 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 2
- 230000003196 chaotropic effect Effects 0.000 claims 2
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 239000012149 elution buffer Substances 0.000 claims 2
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical group [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims 2
- 239000000203 mixture Substances 0.000 claims 2
- 239000005388 borosilicate glass Substances 0.000 claims 1
- 238000005119 centrifugation Methods 0.000 claims 1
- 239000002738 chelating agent Substances 0.000 claims 1
- 238000010828 elution Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 239000008188 pellet Substances 0.000 claims 1
- 239000000377 silicon dioxide Substances 0.000 claims 1
- 239000004094 surface-active agent Substances 0.000 claims 1
- 238000003260 vortexing Methods 0.000 claims 1
Classifications
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- 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
- 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
-
- 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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- 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
- C12Q2523/00—Reactions characterised by treatment of reaction samples
- C12Q2523/30—Characterised by physical treatment
- C12Q2523/32—Centrifugation
-
- 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
- C12Q2527/00—Reactions demanding special reaction conditions
- C12Q2527/125—Specific component of sample, medium or buffer
-
- 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
- C12Q2527/00—Reactions demanding special reaction conditions
- C12Q2527/137—Concentration of a component of medium
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Saccharide Compounds (AREA)
Claims (11)
1. Postupak izoliranja nukleinskih kiselina koje sadrže DNK iz biološkog materijala, postupak koji obuhvata korake:
(i) vršenja liziranja biološkog materijala vodenim rastvorom koji sadrži litij u koncentraciji od 0.05 do manje od 1M, kelatni agens, i surfaktant za proizvodnju vodene lizirane kompozicije,
(ii) tretiranja lizirane kompozicije čvrstim nosačem u prisustvu otopljenog kaotropnog agensa u koncentraciji od 0.05 do 2M i 25% do 60% po zapremini u tekućini otopljenog C1-3 alkanola, pri čemu je navedeni čvrst nosač sposoban za imobilizaciju DNK,
(iii) odvajanja čvrstog nosača od tekućine,
(iv) tretiranja čvrstog nosača prvom otopinom za ispiranje koji sadrži litij u koncentraciji od 0.05 do 1M, pri čemu navedena prva otopina za ispiranje sadrži 20% do 90% po zapremini otopljenog C1-3 alkanola,
(v) odvajanja čvrstog nosača od prve tekućine za ispiranje,
(vi) tretiranja čvrstog nosača drugom tekućinom za ispiranje koja sadrži najmanje 80% po zapremini C1-3 alkanola, pri čemu je ravnoteža ako je prisutna voda,
(vii) odvajanja čvrstog nosača od druge otopine za ispiranje, i
(viii) eluiranja DNK sa čvrstog nosača.
2. Postupak prema patentnom zahtjevu 1 gdje vodena otopina opisana u koraku (i) sadrži litij u koncentraciji od 0.05 do 0.9M.
3. Postupak prema patentnom zahtjevu 1 gdje se otopina za lizu suštinski sastoji od vode, litij klorida u koncentraciji od 0.7 do 0.9M, EDTA ili natrijeve soli istog u koncentraciji od 5 do 15mM i 0.9 do 1.1% po masi SDS, opcionalno gdje u koraku (ii) kaotropni agens je gvanidinij klorid u koncentraciji od 0.6 do 0.7M i alkanol je etanol prisutan u količini od 35-40% po zapremini tekućine.
4. Postupak prema patentnom zahtjevu 1 gdje se otopina za lizu sastoji suštinski od vode, litij klorida u koncentraciji od 0.2 do 0.3M, EDTA ili natrijeve soli istog u koncentraciji od 1 do 5mM, gvanidinij klorida u koncentraciji od 0.6 do 0.7M, SDS u količini od 0.2 do 0.4% po masi i 35-40% po zapremini etanola.
5. Postupak prema bilo kojem od patentnih zahtjeva 1 do 4 gdje se prva tekućina za ispiranje suštinski sastoji od a) vode, litij klorida u koncentraciji od 0.3 do 0.6M, i 45% do 55% po zapremini etanola; ili b) vode, litij klorida u koncentraciji od 0.6 do 1M, i 45% do 55% po zapremini etanola.
6. Postupak prema bilo kojem od patentnih zahtjeva 1 do 5 gdje a) druga tekućina za ispiranje se suštinski sastoji od 80% do 95% po zapremini etanola i vode i/ili b) čvrsti nosač sadrži silicij-dioksid; i/ili c) korak (iii) se vrši u epruveti za centrifugu u kojoj navedeni čvrst nosač je filter i koraci (iii), (v), (vii) i (viii) se vrše uz centrifugiranje.
7. Postupak prema patentnom zahtjevu 6 (c) gdje filter sadrži vlakna borosilikatnog stakla, opcionalno gdje je veliki broj navedenih filtera osiguran u epruveti za centrifugu, opcionalno dalje gdje su četiri filtera osigurana.
8. Postupak prema bilo kojem od patentnih zahtjeva 1 do 7 a) koji se vrši na temperaturi okoline; i/ili b) koji se vrši bez vorteksiranja; i/ili c) gdje biološki materijal iz kojeg će nukleinske kiseline koje sadrže DNK biti izolirane sadrži stanice, opcionalno gdje su stanice u obliku peleta.
9. Kit za ekstrakciju nukleinskih kiselina koje sadrže DNK iz stanica, pri čemu komplet sadrži u kombinaciji:
(A)
(i) vodenu otopina koji sadrži, sastoji se u suštini od, ili se sastoji od vode, litij klorida u koncentraciji od 0.7 do 0.9 mola po litru, EDTA ili natrijeve soli istog u koncentraciji od 5 do 15mM, i 0.9 do 1.1% po masi SDS,
(ii) vodene otopine soli gvanidinija, (poželjno oko 2M),
(iii) etanol,
(iv) prvu tekućinu za ispiranje koja sadrži litij klorid u koncentraciji od 0.3 do 0.6M, pri čemu navedena prva tekućina sadrži etanol u količini od 45% do 55% po zapremini,
(v) drugu tekućini za ispiranje koja sadrži barem 80% po zapremini etanola, pri čemu je ravnoteža ako je prisutna voda,
(vi) elucijski pufer, i
(vii) spin kolonu koja ima filter za imobilizaciju DNK;
ILI
(B)
(i) vodenu otopina koji sadrži, sastoji se u suštini od, ili se sastoji od litij klorida u koncentraciji od 0.2 do 0.3 mola po litru, EDTA ili natrijeve soli istog u koncentraciji od 1 do 5 milimola po litru, SDS u količini od 0.2-0.4% po masi, soli gvanidinija koja ima koncentraciju od 0.6 do 0.7M i etanola u količini od 35-45% po zapremini,
(ii) prvu tekućinu za ispiranje koja sadrži litij klorid u koncentraciji od 0.3 do 0.6M, pri čemu navedena prva tekućina sadrži etanol u količini od 45% do 55% po zapremini,
(iii) drugu tekućinu za ispiranje koja sadrži barem 80% po zapremini etanola, pri čemu je ravnoteža ako je prisutna voda,
(iv) elucijski pufer, i
(v) spin kolonu koja ima filter za imobilizaciju DNK.
10. Komplet prema patentnom zahtjevu 9 gdje filter sadrži barem jednu ploču boro-silikatnih vlakana.
11. Komplet prema patentnom zahtjevu 10 gdje filter sadrži četiri ploče boro-silikatnih vlakana.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB201504459A GB201504459D0 (en) | 2015-03-17 | 2015-03-17 | Isolation of DNA |
PCT/GB2016/050738 WO2016147004A1 (en) | 2015-03-17 | 2016-03-17 | Isolation of nucleic acids |
EP16719888.6A EP3277809B1 (en) | 2015-03-17 | 2016-03-17 | Isolation of nucleic acids |
Publications (1)
Publication Number | Publication Date |
---|---|
HRP20201452T1 true HRP20201452T1 (hr) | 2020-12-25 |
Family
ID=53016238
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
HRP20201452TT HRP20201452T1 (hr) | 2015-03-17 | 2020-09-10 | Izoliranje nukleinskih kiselina |
Country Status (17)
Country | Link |
---|---|
US (1) | US10934539B2 (hr) |
EP (1) | EP3277809B1 (hr) |
JP (1) | JP6713007B2 (hr) |
CN (1) | CN107743521B (hr) |
AU (1) | AU2016231944B2 (hr) |
CY (1) | CY1123363T1 (hr) |
DK (1) | DK3277809T3 (hr) |
ES (1) | ES2819903T3 (hr) |
GB (1) | GB201504459D0 (hr) |
HR (1) | HRP20201452T1 (hr) |
HU (1) | HUE051568T2 (hr) |
LT (1) | LT3277809T (hr) |
PL (1) | PL3277809T3 (hr) |
PT (1) | PT3277809T (hr) |
RS (1) | RS60850B1 (hr) |
SI (1) | SI3277809T1 (hr) |
WO (1) | WO2016147004A1 (hr) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11015186B2 (en) | 2016-09-27 | 2021-05-25 | Abbott Molecular Inc. | Maximizing DNA yield of blood specimens collected in rapid clot tubes |
CN108427000B (zh) * | 2017-02-15 | 2021-06-08 | 广州市锐博生物科技有限公司 | 一种捕获核酸结合蛋白的方法和试剂盒 |
GB2581489B (en) | 2019-02-15 | 2021-02-24 | Revolugen Ltd | Purification method |
DE102020135124A1 (de) | 2020-12-30 | 2022-06-30 | Aj Innuscreen Gmbh | Verfahren und System zur schnellen Isolierung von Nukleinsäuren direkt aus Vollblutproben |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE210671T1 (de) | 1994-02-11 | 2001-12-15 | Qiagen Gmbh | Verfahren zur trennung von doppelstrang/einzelstrangnukleinsäurestrukturen |
GB9425138D0 (en) * | 1994-12-12 | 1995-02-08 | Dynal As | Isolation of nucleic acid |
JP2002516663A (ja) * | 1998-04-02 | 2002-06-11 | リー キュンジル | ガラス微細繊維コラムとこれを利用したプラスミドDNA製造および精製方法(GlassmicrofibercolumnandmethodforthepreparationandpurificationofplasmidDNAusingthesame) |
US7148343B2 (en) | 2001-10-12 | 2006-12-12 | Gentra Systems, Inc. | Compositions and methods for using a solid support to purify RNA |
US20050032105A1 (en) * | 2001-10-12 | 2005-02-10 | Bair Robert Jackson | Compositions and methods for using a solid support to purify DNA |
EP1529840A1 (en) * | 2003-11-04 | 2005-05-11 | Qiagen GmbH | A rapid and low cost method for isolating nucleic acid |
US20060099605A1 (en) * | 2004-11-11 | 2006-05-11 | Hall Gerald E Jr | Devices and methods for isolating RNA |
US20070042384A1 (en) | 2004-12-01 | 2007-02-22 | Weiwei Li | Method for isolating and modifying DNA from blood and body fluids |
DE102006019650A1 (de) | 2006-04-25 | 2007-10-31 | InViTek Gesellschaft für Biotechnik & Biodesign mbH | Formulierungen und Verfahren zur Isolierung von Nukleinsäuren aus beliebigen komplexen Ausgangsmaterialien und nachfolgende komplexe Genanalytik |
EP3617321A3 (en) | 2006-05-31 | 2020-04-29 | Sequenom, Inc. | Methods and compositions for the extraction and amplification of nucleic acid from a sample |
DK2521780T3 (en) | 2010-01-07 | 2017-12-04 | Bigtec Private Ltd | Process for Isolation of Nucleic Acids and Kits thereof |
DK2539449T3 (en) | 2010-02-26 | 2018-08-06 | Qiagen Gmbh | PROCEDURE FOR PARALLEL ISOLATION AND CLEANING RNA AND DNA |
JP5714291B2 (ja) | 2010-10-15 | 2015-05-07 | 独立行政法人農業・食品産業技術総合研究機構 | 抗酸菌dnaの抽出精製法 |
GB201415674D0 (en) | 2014-09-04 | 2014-10-22 | Moorlodge Biotech Ventures Ltd | Nucleic acid analysis |
-
2015
- 2015-03-17 GB GB201504459A patent/GB201504459D0/en not_active Ceased
-
2016
- 2016-03-17 ES ES16719888T patent/ES2819903T3/es active Active
- 2016-03-17 LT LTEP16719888.6T patent/LT3277809T/lt unknown
- 2016-03-17 WO PCT/GB2016/050738 patent/WO2016147004A1/en active Application Filing
- 2016-03-17 AU AU2016231944A patent/AU2016231944B2/en active Active
- 2016-03-17 PL PL16719888T patent/PL3277809T3/pl unknown
- 2016-03-17 PT PT167198886T patent/PT3277809T/pt unknown
- 2016-03-17 US US15/558,864 patent/US10934539B2/en active Active
- 2016-03-17 SI SI201630924T patent/SI3277809T1/sl unknown
- 2016-03-17 CN CN201680028490.3A patent/CN107743521B/zh active Active
- 2016-03-17 HU HUE16719888A patent/HUE051568T2/hu unknown
- 2016-03-17 JP JP2017567557A patent/JP6713007B2/ja active Active
- 2016-03-17 DK DK16719888.6T patent/DK3277809T3/da active
- 2016-03-17 RS RS20201088A patent/RS60850B1/sr unknown
- 2016-03-17 EP EP16719888.6A patent/EP3277809B1/en active Active
-
2020
- 2020-09-10 HR HRP20201452TT patent/HRP20201452T1/hr unknown
- 2020-09-23 CY CY20201100898T patent/CY1123363T1/el unknown
Also Published As
Publication number | Publication date |
---|---|
EP3277809A1 (en) | 2018-02-07 |
PT3277809T (pt) | 2020-09-23 |
HUE051568T2 (hu) | 2021-03-01 |
JP6713007B2 (ja) | 2020-06-24 |
LT3277809T (lt) | 2020-11-10 |
ES2819903T3 (es) | 2021-04-19 |
RS60850B1 (sr) | 2020-10-30 |
US20180066246A1 (en) | 2018-03-08 |
AU2016231944B2 (en) | 2021-07-29 |
AU2016231944A8 (en) | 2017-12-07 |
US10934539B2 (en) | 2021-03-02 |
PL3277809T3 (pl) | 2020-11-16 |
SI3277809T1 (sl) | 2020-11-30 |
CN107743521A (zh) | 2018-02-27 |
GB201504459D0 (en) | 2015-04-29 |
WO2016147004A1 (en) | 2016-09-22 |
AU2016231944A1 (en) | 2017-11-09 |
JP2018508239A (ja) | 2018-03-29 |
DK3277809T3 (da) | 2020-09-28 |
CN107743521B (zh) | 2021-01-08 |
EP3277809B1 (en) | 2020-06-24 |
CY1123363T1 (el) | 2021-12-31 |
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