JP3860174B2 - 同時多検体ハイブリダイゼーション方法 - Google Patents
同時多検体ハイブリダイゼーション方法 Download PDFInfo
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- JP3860174B2 JP3860174B2 JP2004056511A JP2004056511A JP3860174B2 JP 3860174 B2 JP3860174 B2 JP 3860174B2 JP 2004056511 A JP2004056511 A JP 2004056511A JP 2004056511 A JP2004056511 A JP 2004056511A JP 3860174 B2 JP3860174 B2 JP 3860174B2
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- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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- 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
- C12Q2527/109—Pressure
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- 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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- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
1×106個のHeLa細胞より、RNeasy Mini Kit(Qiagen社製)を用いて10μgのTotal RNAを調製した。調製した10μgのHeLa Total RNAから、Labelstar Array Kit(Qiagen社製)とCy3−dUTP(Amersham Biosciences社製)を用いてCy3標識HeLa cDNAを調製し、これをサンプル生体高分子とした。サンプル生体高分子を0.5%SDSを含有する5×SSC溶液を用いて、液量120μLに調製した(サンプル生体高分子を含む溶液の溶質モル濃度:1.67mol/L)。
保湿液として、0.5%SDSを含有する4.5×SSCを600μL(保湿液の溶質モル濃度:1.50mol/L、サンプル生体高分子を含む溶液との溶質モル濃度の差異:−10%)を用いた以外は、実施例1と同様にして、ハイブリダイゼーションを行なった。ハイブリダイズ後、サンプル生体高分子を含む溶液の液量を測定したところ、滴下した時点よりも4.9μL増加しており、液量の変化は4.1%であった。このように、サンプル生体高分子を含む溶液の体積が増加して当該溶液が親水性領域からあふれてしまうことなく、サンプル生体高分子を含む溶液がプローブ生体高分子が固定されているスライドグラスのみに接触した状態で、ハイブリダイゼーションを完了することができた。
保湿液として、0.5%SDSを含有する5.4×SSCを600μL(保湿液の溶質モル濃度:1.80mol/L、サンプル生体高分子を含む溶液との溶質モル濃度の差異:+8%)を用いた以外は、実施例1と同様にして、ハイブリダイゼーションを行なった。ハイブリダイズ後、サンプル生体高分子を含む溶液の液量を測定したところ、滴下した時点よりも12.0μL減少しており、液量の変化は10.0%であった。このように、サンプル生体高分子を含む溶液がマイクロアレイ上で乾燥してしまうことなく、サンプル生体高分子を含む溶液がプローブ生体高分子が固定されているスライドグラスのみに接触した状態で、ハイブリダイゼーションを完了することができた。
保湿液として、0.5%SDSを含有する4×SSCを600μL(保湿液の溶質モル濃度:1.33mol/L、サンプル生体高分子を含む溶液との溶質モル濃度の差異:−20%)を用いた以外は、実施例1と同様にして、ハイブリダイゼーションを行なった。ハイブリダイズ後、サンプル生体高分子を含む溶液の液量を測定したところ、滴下した時点よりも17.4μL増加しており(液量の変化:14.5%)、サンプル生体高分子を含む溶液の著しい液量増加により、当該溶液があふれてしまって、正常にハイブリダイゼーションが行なえていなかった。
保湿液として、0.5%SDSを含有する5.5×SSCを600μL(保湿液の溶質モル濃度:1.83mol/L、サンプル生体高分子を含む溶液との溶質モル濃度の差異:+10%)を用いた以外は、実施例1と同様にして、ハイブリダイゼーションを行なった。ハイブリダイズ後、サンプル生体高分子を含む溶液の液量を測定したところ、滴下した時点よりも14.3μL減少しており(液量の変化:11.9%)、ハイブリダイゼーション中の蒸気拡散によりサンプル生体高分子を含む溶液の著しい液量減が起こり、正常にハイブリダイゼーションが行なえていなかった。
保湿液として、0.5%SDSを含有する6×SSCを600μL(保湿液の溶質モル濃度:2.00mol/L、サンプル生体高分子を含む溶液との溶質モル濃度の差異:+20%)を用いた以外は、実施例1と同様にして、ハイブリダイゼーションを行なった。ハイブリダイズ後、サンプル生体高分子を含む溶液の液量を測定したところ、滴下した時点よりも21.9μL減少しており(液量の変化:18.2%)、ハイブリダイゼーション中の蒸気拡散によりサンプル生体高分子を含む溶液の著しい液量減が起こり、正常にハイブリダイゼーションが行なえていなかった。
Claims (1)
- 1枚のスライドグラス上に、複数種のプローブ生体高分子がスポットされ固定されている親水性領域が複数配列されてなり、この複数配列された親水性領域の周りにプローブ生体高分子が固定されていない疎水性領域が形成されたマイクロアレイを用いて、多検体について同時にハイブリダイゼーションを行う方法であって、
サンプル生体高分子を含む溶液がスライドグラス上の親水性領域に接触した状態で、サンプル生体高分子を含む溶液と同一の蒸気圧を有する溶液を入れた密閉容器中でサンプル生体高分子とプローブ生体高分子とのハイブリダイゼーションを行なうことを特徴とする同時多検体ハイブリダイゼーション方法。
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004056511A JP3860174B2 (ja) | 2004-03-01 | 2004-03-01 | 同時多検体ハイブリダイゼーション方法 |
PCT/JP2005/003801 WO2005083119A2 (en) | 2004-03-01 | 2005-02-25 | Hybridization method as well as hybridization microarray and hybridization kit |
KR1020067018699A KR100827584B1 (ko) | 2004-03-01 | 2005-02-25 | 혼성화 방법 및 혼성화용 마이크로어레이 및 혼성화용 키트 |
DE602005010556T DE602005010556D1 (de) | 2004-03-01 | 2005-02-25 | Hybridisierungsverfahren sowie hybridisierungs-mikroarray und hybridisierungskit |
CN2005800061983A CN1926246B (zh) | 2004-03-01 | 2005-02-25 | 杂交方法、杂交微阵列和杂交试剂盒 |
US10/589,497 US20080312100A1 (en) | 2004-03-01 | 2005-02-25 | Hybridization Method as Well as Hybridization Microarray and Hybridization Kit |
EP05720073A EP1721009B1 (en) | 2004-03-01 | 2005-02-25 | Hybridization method as well as hybridization microarray and hybridization kit |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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JP2004056511A JP3860174B2 (ja) | 2004-03-01 | 2004-03-01 | 同時多検体ハイブリダイゼーション方法 |
Publications (2)
Publication Number | Publication Date |
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JP2005249421A JP2005249421A (ja) | 2005-09-15 |
JP3860174B2 true JP3860174B2 (ja) | 2006-12-20 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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JP2004056511A Expired - Fee Related JP3860174B2 (ja) | 2004-03-01 | 2004-03-01 | 同時多検体ハイブリダイゼーション方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20080312100A1 (ja) |
EP (1) | EP1721009B1 (ja) |
JP (1) | JP3860174B2 (ja) |
KR (1) | KR100827584B1 (ja) |
CN (1) | CN1926246B (ja) |
DE (1) | DE602005010556D1 (ja) |
WO (1) | WO2005083119A2 (ja) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4779514B2 (ja) * | 2005-09-02 | 2011-09-28 | ニプロ株式会社 | 試験片およびその製造方法 |
WO2007063719A1 (ja) * | 2005-11-29 | 2007-06-07 | Nec Corporation | 電気泳動チップ、電気泳動装置、および試料分析方法 |
US20070161119A1 (en) * | 2006-01-12 | 2007-07-12 | Peck Bill J | Methods for fabricating chemical arrays |
WO2009058867A2 (en) * | 2007-10-29 | 2009-05-07 | Primorigen Biosciences, Llc | Affinity measurements using frameless multiplexed microarrays |
CA2723707C (en) * | 2008-05-09 | 2016-08-23 | Akonni Biosystems | Microarray system |
CN104614516B (zh) * | 2014-01-09 | 2016-03-30 | 南京医科大学第一附属医院 | 确保实验结果好的载玻片孵育器 |
CN103743895B (zh) * | 2014-01-09 | 2015-05-27 | 南通大学 | 可拆卸的载玻片孵育器 |
USD815752S1 (en) * | 2014-11-28 | 2018-04-17 | Randox Laboratories Ltd. | Biochip well |
CN107904283A (zh) * | 2017-11-09 | 2018-04-13 | 武汉康录生物技术股份有限公司 | 一种即用型多探针杂交玻片及其制备方法和应用 |
CN108728346B (zh) * | 2018-08-20 | 2024-06-18 | 中山康源基因技术科技有限公司 | 间歇式重力微流芯片杂交方法 |
Family Cites Families (11)
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US6589726B1 (en) * | 1991-09-04 | 2003-07-08 | Metrigen, Inc. | Method and apparatus for in situ synthesis on a solid support |
US6555361B1 (en) * | 1999-03-24 | 2003-04-29 | Corning Incorporated | Hybridization chamber for high density nucleic acid arrays |
US6773676B2 (en) * | 1999-04-27 | 2004-08-10 | Agilent Technologies, Inc. | Devices for performing array hybridization assays and methods of using the same |
CN1356392A (zh) * | 2000-12-01 | 2002-07-03 | 吴昌 | 一种核酸分子杂交方法及所用的装置 |
JP2002267667A (ja) * | 2001-03-08 | 2002-09-18 | Hitachi Software Eng Co Ltd | マイクロアレイ及びマイクロアレイ用基板 |
CA2450676C (en) * | 2001-03-09 | 2010-03-30 | Biomicro Systems, Inc. | Method and system for microfluidic interfacing to arrays |
US6558907B2 (en) * | 2001-05-16 | 2003-05-06 | Corning Incorporated | Methods and compositions for arraying nucleic acids onto a solid support |
US20050255466A1 (en) * | 2002-03-28 | 2005-11-17 | Deutsches Krebsforschungszntrum | Method and system for determining absolute mrna quantities |
US20030194709A1 (en) * | 2002-04-10 | 2003-10-16 | Xing Yang | Hydrophobic zone device |
US7498176B2 (en) * | 2002-09-27 | 2009-03-03 | Roche Nimblegen, Inc. | Microarray with hydrophobic barriers |
US20040152083A1 (en) * | 2003-01-31 | 2004-08-05 | Leproust Eric M. | Multiple arrays with surface energy transition to maintain separation of samples on the arrays |
-
2004
- 2004-03-01 JP JP2004056511A patent/JP3860174B2/ja not_active Expired - Fee Related
-
2005
- 2005-02-25 US US10/589,497 patent/US20080312100A1/en not_active Abandoned
- 2005-02-25 KR KR1020067018699A patent/KR100827584B1/ko not_active IP Right Cessation
- 2005-02-25 CN CN2005800061983A patent/CN1926246B/zh not_active Expired - Fee Related
- 2005-02-25 DE DE602005010556T patent/DE602005010556D1/de active Active
- 2005-02-25 EP EP05720073A patent/EP1721009B1/en not_active Expired - Fee Related
- 2005-02-25 WO PCT/JP2005/003801 patent/WO2005083119A2/en active Application Filing
Also Published As
Publication number | Publication date |
---|---|
DE602005010556D1 (de) | 2008-12-04 |
KR20060122954A (ko) | 2006-11-30 |
CN1926246A (zh) | 2007-03-07 |
WO2005083119A2 (en) | 2005-09-09 |
EP1721009B1 (en) | 2008-10-22 |
CN1926246B (zh) | 2011-07-06 |
KR100827584B1 (ko) | 2008-05-07 |
WO2005083119A3 (en) | 2005-10-27 |
EP1721009A2 (en) | 2006-11-15 |
US20080312100A1 (en) | 2008-12-18 |
JP2005249421A (ja) | 2005-09-15 |
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