JP2008500836A5 - - Google Patents
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- JP2008500836A5 JP2008500836A5 JP2007515379A JP2007515379A JP2008500836A5 JP 2008500836 A5 JP2008500836 A5 JP 2008500836A5 JP 2007515379 A JP2007515379 A JP 2007515379A JP 2007515379 A JP2007515379 A JP 2007515379A JP 2008500836 A5 JP2008500836 A5 JP 2008500836A5
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本発明のシステムおよび方法は、一部の実施形態ではマイクロスフィアすなわちビーズなどのマイクロキャリア要素を含むマイクロエマルジョン液滴すなわちボーラスなどのマイクロリアクタ要素を用いる。マイクロキャリア要素は、ボーラスすなわちマイクロエマルジョン液滴の中に含まれており、DNA産物またはRNA産物を捕捉するのに有用である。すなわち、増幅されたDNAまたはRNAを、マイクロキャリア要素に対して化学的に結合させることができる。あるいは、増幅されたDNAまたはRNAは、マイクロキャリア要素を利用せずに、マイクロリアクタ要素によって運ばれてもよい。例えば、ボーラスすなわちマイクロエマルジョン液滴の中でPCR反応を実行してよく、その後、結果として得られたボーラスすなわちマイクロエマルジョン液滴を、次の処理工程に送ってよい。
Claims (27)
- 微細加工構造であって、
1つの熱循環チャンバ内に1つだけのマイクロキャリア要素が入るように、クローン配列決定テンプレートの複数の複製物を運ぶマイクロキャリア要素を複数の熱循環チャンバに分配する分配流路を備え、
前記熱循環チャンバは、前記クローン配列決定テンプレートから熱循環伸長断片を生成するように構成され、
前記微細加工構造は、さらに、
前記熱循環チャンバに接続され、前記伸長断片を捕捉および濃縮する精製チャンバと、
前記精製チャンバに接続され、前記伸長断片を分析する成分分離流路と、を備える、微細加工構造。 - 請求項1に記載の微細加工構造であって、前記マイクロキャリア要素は、前記クローン配列決定テンプレートの前記複数の複製物を運ぶ、微細加工構造。
- 請求項1に記載の微細加工構造であって、前記マイクロキャリア要素は、マイクロスフィアすなわちビーズである、微細加工構造。
- 請求項1に記載の微細加工構造であって、前記配列決定テンプレートは、DNAまたはRNAの配列決定テンプレートである、微細加工構造。
- 配列決定を実行するためのシステムであって、
各々が配列決定テンプレートの複数のクローン複製物を運ぶ複数のマイクロキャリア要素を、複数の熱循環チャンバに分配するマイクロ流体分配手段であって、前記分配手段と前記複数の熱循環チャンバとは微細加工構造に集積されている、マイクロ流体分配手段と、
1つの熱循環チャンバ内に1つだけのマイクロキャリア要素が流れ込むことを保証する手段と、
前記熱循環チャンバを備え、前記配列決定テンプレートから熱循環伸長断片を生成する伸長手段と、
前記伸長断片を捕捉、精製、および、濃縮する精製チャンバ手段と、
前記伸長断片を分析する成分分離手段と、を備える、システム。 - 請求項5に記載のシステムであって、前記マイクロキャリア要素は、マイクロスフィアすなわちビーズである、システム。
- 請求項5に記載のシステムであって、さらに、少なくともいくつかは1つのマイクロキャリア要素を含む複数のマイクロリアクタ要素を備える、システム。
- 請求項7に記載のシステムであって、前記複数のマイクロリアクタ要素のそれぞれは、ボーラスすなわちマイクロエマルジョン液滴を備える、システム。
- 微細加工構造であって、
クローン配列決定テンプレートの複数の複製物を運ぶ複数のマイクロスフィアを複数の熱循環チャンバに分配する分配流路と、
熱循環チャンバの出口ポートに配置され、1つの熱循環チャンバ内に1つだけのマイクロスフィアが流れ込むことを保証する自動弁と、を備え、
前記熱循環チャンバは、前記クローン配列決定テンプレートから伸長断片を生成するように構成され、
前記微細加工構造は、さらに、
前記熱循環チャンバに接続され、前記伸長断片を捕捉および濃縮する精製チャンバと、
前記精製チャンバに接続され、前記伸長断片を分析する成分分離流路と、を備える、微細加工構造。 - 請求項9に記載の微細加工構造であって、マイクロスフィアの直径は、1から100ミクロンの間である、微細加工構造。
- 請求項10に記載の微細加工構造であって、マイクロスフィアの直径は、10ミクロンである、微細加工構造。
- 請求項9に記載の微細加工構造であって、各熱循環チャンバと、分離流路との間で、流体が行き来するようになっている、微細加工構造。
- 請求項9に記載の微細加工構造であって、前記配列決定テンプレートは、DNAまたはRNAの配列決定テンプレートである、微細加工構造。
- 微細加工構造であって、
配列決定テンプレートを運ぶマイクロスフィアを熱循環チャンバに分配するマイクロ流体分配流路手段と、
1つの熱循環チャンバに1つだけのマイクロスフィアが流れ込むことを保証する自動弁手段と、
前記熱循環チャンバを備え、配列決定テンプレートから熱循環伸長断片を生成する伸長手段と、
前記伸長断片を捕捉、精製、および、濃縮する集積精製チャンバ手段と、
前記伸長断片を分析する成分分離手段と、を備える、微細加工構造。 - 請求項14に記載の微細加工構造であって、前記伸長手段は、複数の熱循環チャンバを備えるサンガー伸長手段である、微細加工構造。
- 請求項15に記載の微細加工構造であって、前記自動弁手段は、前記熱循環チャンバの出口ポートの自動弁を含む、微細加工構造。
- 請求項16に記載の微細加工構造であって、前記精製チャンバ手段は、前記熱循環チャンバに接続された精製チャンバを備える、微細加工構造。
- 請求項17に記載の微細加工構造であって、前記成分分離手段は、前記精製チャンバに接続された複数のマイクロ流路を備えるキャピラリアレイ電気泳動手段である、微細加工構造。
- 請求項14に記載の微細加工構造であって、前記配列決定テンプレートは、DNAまたはRNAの配列決定テンプレートである、微細加工構造。
- 配列決定を実行するためのシステムであって、
DNAまたはRNAを断片にせん断する手段と、
前記断片をライゲーションさせて、所望の環状産物と不純物である線形産物との混合物を形成する手段と、
前記不純物である線形産物を選択的に除去する手段と、
1つの配列決定テンプレートの複数のクローン複製物を運ぶマイクロスフィアを生成する手段と、
配列決定テンプレートを有するマイクロスフィアを選択する手段と、
配列決定テンプレートを有する選択されたマイクロスフィアを熱循環チャンバに分配するマイクロ流体分配流路手段と、
統計的に1つの熱循環チャンバに1つだけのマイクロスフィアが流れ込むことを保証する手段と、
前記熱循環チャンバを備え、前記配列決定テンプレートから熱循環伸長断片を生成する伸長手段と、
前記伸長断片を捕捉、精製、および、濃縮する精製チャンバ手段と、
前記伸長断片を分析する成分分離手段と、を備える、システム。 - 請求項20に記載のシステムであって、前記保証手段は、前記熱循環チャンバに配置された自動弁と、光学検出器と、タイミング機構との内の少なくとも1つである、システム。
- 請求項21に記載のシステムであって、前記光学検出器は、マイクロスフィアからの散乱光を検出する光学スキャナである、システム。
- 請求項20に記載のシステムであって、前記タイミング機構は、熱循環チャンバの流入口に隣接して配置された弁を備える、システム。
- DNA配列決定を実行するためのシステムであって、
DNAをDNA断片にせん断する手段と、
前記DNA断片をライゲーションさせて、所望の環状産物と不純物である線形産物との混合物を形成する手段と、
前記不純物である線形産物を選択的に除去するエキソヌクレアーゼ分解手段と、
1つのDNA配列決定テンプレートの複数のクローン複製物を運ぶマイクロスフィアを生成するエマルジョンPCR反応手段と、
DNA配列決定テンプレートを有するマイクロスフィアを選択する蛍光活性化細胞選別(FACS)手段と、
DNA配列決定テンプレートを有する選択されたマイクロスフィアを熱循環チャンバに分配するマイクロ流体分配流路手段と、
統計的に1つの熱循環チャンバに1つだけのマイクロスフィアが流れ込むことを保証する自動弁手段と、
前記熱循環チャンバを備え、前記DNA配列決定テンプレートの複数の複製物を運ぶ前記マイクロスフィアから熱循環伸長断片を生成するサンガー伸長手段と、
前記伸長断片を捕捉、精製、および、濃縮する集積精製チャンバ手段と、
前記伸長断片を分析するキャピラリアレイ電気泳動手段と、を備える、システム。 - 微細加工構造に集積された分配流路と複数の第2流路とを備える前記微細加工構造であって、
(a)前記第2流路のそれぞれは、前記分配流路に接続されており、
(b)前記分配流路は、各々が配列決定テンプレートの複数のクローン複製物を運ぶ複数のマイクロキャリア要素を、複数の第2流路に分配するように構成されており、
(c)前記複数の第2流路のそれぞれは、成分分離流路に接続された精製チャンバに接続された熱循環リアクタを備え、
(d)前記熱循環リアクタのそれぞれは、その中に配置されたマイクロキャリア要素から熱循環伸長断片を生成するように構成されており、前記精製チャンバは、前記伸長断片を捕捉および濃縮するように構成されており、前記成分分離流路は、前記伸長断片を、それらの相対的なサイズに基づいて、互いに分離するように構成されており、
(e)前記微細加工構造は、前記分配流路からの前記複数のマイクロキャリア要素の分配の間に1つだけのマイクロキャリア要素が個々の第2流路内に入り得るように、構成されている、
微細加工構造。 - 請求項25に記載の微細加工構造であって、さらに、
前記分配流路内のマイクロキャリア要素の位置を決定するためのタイミング機構と光学検出器とのうちの少なくとも1つを備える、
微細加工構造。 - 請求項25に記載の微細加工構造であって、さらに、
まだマイクロキャリア要素を含んでいない熱循環リアクタにマイクロキャリア要素が流れ込むことを許容し、既にマイクロキャリア要素を含む熱循環リアクタにマイクロキャリア要素が流れ込むことを防止する弁とタイミング機構とを備える、
微細加工構造。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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US57610204P | 2004-06-01 | 2004-06-01 | |
US11/139,018 US7799553B2 (en) | 2004-06-01 | 2005-05-25 | Microfabricated integrated DNA analysis system |
PCT/US2005/018678 WO2005118867A2 (en) | 2004-06-01 | 2005-05-26 | Microfabricated integrated dna analysis system |
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JP2008500836A JP2008500836A (ja) | 2008-01-17 |
JP2008500836A5 true JP2008500836A5 (ja) | 2011-10-20 |
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JP2007515379A Pending JP2008500836A (ja) | 2004-06-01 | 2005-05-26 | 微細加工集積dna分析システム |
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US (3) | US7799553B2 (ja) |
EP (1) | EP1774042A4 (ja) |
JP (1) | JP2008500836A (ja) |
KR (1) | KR101214270B1 (ja) |
WO (1) | WO2005118867A2 (ja) |
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KR100960066B1 (ko) | 2008-05-14 | 2010-05-31 | 삼성전자주식회사 | 동결건조시약이 저장된 미세유동장치 및 이를 이용한시료분석방법 |
US8323568B2 (en) | 2008-06-13 | 2012-12-04 | Honeywell International Inc. | Magnetic bead assisted sample conditioning system |
JP5311518B2 (ja) | 2008-10-06 | 2013-10-09 | コーニンクレッカ フィリップス エヌ ヴェ | マイクロ流体装置 |
CN102341691A (zh) | 2008-12-31 | 2012-02-01 | 尹特根埃克斯有限公司 | 具有微流体芯片的仪器 |
EP2438154A1 (en) | 2009-06-02 | 2012-04-11 | Integenx Inc. | Fluidic devices with diaphragm valves |
CN102803147B (zh) * | 2009-06-05 | 2015-11-25 | 尹特根埃克斯有限公司 | 通用样品准备系统以及在一体化分析系统中的用途 |
-
2005
- 2005-05-25 US US11/139,018 patent/US7799553B2/en active Active
- 2005-05-26 WO PCT/US2005/018678 patent/WO2005118867A2/en active Application Filing
- 2005-05-26 JP JP2007515379A patent/JP2008500836A/ja active Pending
- 2005-05-26 KR KR1020067026091A patent/KR101214270B1/ko active IP Right Grant
- 2005-05-26 EP EP05804847A patent/EP1774042A4/en not_active Withdrawn
-
2010
- 2010-07-27 US US12/844,544 patent/US20110020920A1/en not_active Abandoned
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2012
- 2012-02-13 US US13/372,376 patent/US8420318B2/en active Active
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