JP2008500836A - 微細加工集積dna分析システム - Google Patents
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Abstract
【解決手段】ゲノム分析のための方法および装置が提供されている。マイクロ流体分配流路を備える微細加工構造は、配列決定テンプレートの複数の複製物を有するマイクロリアクタ要素を微細加工熱循環チャンバに分配するよう構成されている。マイクロリアクタ要素は、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含んでよい。マイクロキャリア要素は、マイクロスフィアを含んでよい。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ために、熱循環チャンバの出口ポートに配置された自動弁、光学スキャナ、または、タイミング構成を用いてよく、熱循環チャンバ内では、熱循環伸長断片が生成される。伸長産物は、集積オリゴヌクレオチドゲル捕捉チャンバで、捕捉、精製、および、濃縮される。精製された伸長断片は、微細加工成分分離装置を用いて分析される。微細加工構造は、DNAまたはRNAに関する配列決定やその他の遺伝分析を実行する処理で用いられてよい。
【選択図】図9A
Description
本願は、米国特許法第119条(e)に従って、「MICROBEAD INTEGRATED DNA ANALYSIS SYSTEM(MINDS)」と題され、2004年6月1日に出願された米国特許仮出願第60/576,102号(代理人整理番号UCALP054P)からの優先権を主張するものであり、ここに参照によって組み込まれる。
本発明の技術とメカニズムとは、整理番号HG001399,PO1 CA77664,U01AI056472の米国国立衛生研究所の提供(NIH Grant)による政府のサポートによって、なされた。
Claims (44)
- 微細加工構造であって、
1つの熱循環チャンバ内に1つだけのマイクロリアクタ要素が入るように、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リアクタ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、
マイクロリアクタ要素から熱循環伸長断片が生成され、
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、前記マイクロリアクタ要素は、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マイクロキャリア要素を備える、微細加工構造。
- 請求項1に記載の微細加工構造であって、前記マイクロリアクタ要素は、ボーラスすなわちマイクロエマルジョン液滴である、微細加工構造。
- 請求項3に記載の微細加工構造であって、前記マイクロリアクタ要素は、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マイクロスフィアを備える、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リアクタ要素を生成する手段と、
配列決定テンプレートを有するマイクロリアクタ要素を選択する手段と、
配列決定テンプレートを有する選択されたマイクロリアクタ要素を熱循環チャンバに分配す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つだけのマイクロリアクタ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の複数の複製物を運ぶ前記マイクロリアクタ要素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6に記載のシステムであって、前記マイクロリアクタ要素は、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マイクロキャリア要素を備える、システム。
- 請求項6に記載のシステムであって、前記マイクロリアクタ要素は、ボーラスすなわちマイクロエマルジョン液滴である、システム。
- 請求項8に記載のシステムであって、前記マイクロリアクタ要素は、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マイクロスフィアを備える、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、
マイクロスフィアから熱循環伸長断片が生成され、
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、約1から100ミクロンの間である、微細加工構造。
- 請求項11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、約10ミクロンである、微細加工構造。
- 請求項10に記載の微細加工構造であって、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10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を運ぶマイクロスフィア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15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16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17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18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15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。
- 微細加工装置であって、
クローンテンプレートを運ぶマイクロスフィアを受け入れるよう構成され、流入ポートと流出ポートとを有する熱循環チャンバを備え、前記流出ポートは、前記チャンバ内でマイクロスフィアを捕捉して前記熱循環チャンバへのさらなる流入を実質的に遮断するよう構成された狭窄部を備える、微細加工装置。 - 請求項21に記載の微細加工装置であって、前記狭窄部の形状は、実質的に円形である、微細加工装置。
- 請求項21に記載の微細加工装置であって、前記狭窄部の形状は、実質的に半円形である、微細加工装置。
- 請求項21に記載の微細加工装置であって、前記流入ポートとの間で流体が行き来する流入路には、第1の弁が配置されており、前記流出ポートとの間で流体が行き来する流出路には、第2の弁が配置されている、微細加工装置。
- 請求項21に記載の微細加工装置であって、動作中、前記第2の弁が前記第1の弁より前に閉じられることで、熱循環の前に、マイクロスフィアを、前記狭窄部から前記熱循環チャンバの本体部分の中に移動させる、微細加工装置。
- 請求項21に記載の微細加工装置であって、さらに、前記熱循環チャンバの前記流出ポートとの間で流体が行き来する精製チャンバと、成分分離装置との間で流体が行き来する前記精製チャンバの出力ポートとを備える、微細加工装置。
- 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27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27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27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空気入力を備える、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。 - 配列決定を実行するための処理であって、
DNAまたはRNAを断片にせん断する工程と、
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工程と、
前記不純物である線形産物を選択的に除去する工程と、
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リアクタ要素を生成する工程と、
配列決定テンプレートを有するマイクロリアクタ要素を選択する工程と、
前記配列決定テンプレートを有する前記マイクロリアクタ要素を熱循環チャンバ内に分配する工程と、
前記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リアクタ要素から熱循環伸長断片を生成する工程と、
前記伸長断片を捕捉および濃縮する工程と、
前記熱循環伸長断片を分析する工程と、を備える、処理。 - 請求項32に記載の処理であって、前記マイクロリアクタ要素は、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マイクロキャリア要素を備える、処理。
- 請求項32に記載の処理であって、前記マイクロリアクタ要素は、ボーラスすなわちマイクロエマルジョン液滴である、処理。
- 請求項34に記載の処理であって、前記マイクロリアクタ要素は、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マイクロスフィアを備える、処理。
- 請求項32に記載の処理であって、前記分配工程は、1つの熱循環チャンバに1つだけのマイクロリアクタ要素が入るように実行される、処理。
- 請求項36に記載の処理であって、さらに、前記熱循環チャンバの出口ポートに配置された自動弁を用いて、1つの熱循環チャンバに1つだけのマイクロリアクタ要素が流れ込むことを保証する工程を備える、処理。
- 請求項32に記載の処理であって、前記生成工程は、エマルジョンPCR反応またはフロースルー型PCR処理によって、配列決定テンプレートの複数のクローン複製物を生成する工程を備える、処理。
- 請求項32に記載の処理であって、前記選択工程は、蛍光活性化細胞選別(FACS)である、処理。
- DNA配列決定を実行するための処理であって、
DNAをDNA断片にせん断する工程と、
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工程と、
エキソヌクレアーゼ分解によって、前記不純物である線形産物を選択的に除去する工程と、
エマルジョンPCR反応によって、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工程と、
蛍光活性化細胞選別(FACS)によって、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工程と、
前記DNA配列決定テンプレートを有する前記マイクロスフィアを熱循環チャンバに分配する工程と、
熱循環チャンバの出口ポートに配置された自動弁を用いて、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工程と、
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工程と、
前記伸長断片を捕捉、精製、および、濃縮する工程と、
前記熱循環伸長断片を分析する工程と、を備える、処理。 - 請求項40に記載の処理であって、前記捕捉工程は、オリゴヌクレオチド捕捉マトリクスを用いる、処理。
- 配列決定で用いる処理であって、
熱循環チャンバの流入ポートにおいてクローンテンプレートを運ぶマイクロスフィアを受け入れる工程と、
前記チャンバの流出ポートに配置された狭窄部を用いて、前記チャンバ内で前記マイクロスフィアを捕捉して前記チャンバへのさらなる流入を実質的に遮断する工程と、を備える、処理。 - 分析のための方法であって、
配列決定テンプレートを運ぶマイクロスフィアを生成する工程と、
熱循環チャンバの出力ポートに配置された狭窄部を用いて、前記熱循環チャンバ内に前記マイクロスフィアを配置して、前記チャンバへのさらなる流入を実質的に遮断する工程と、を備える、方法。 - 請求項43に記載の方法であって、前記熱循環チャンバの流入ポートに、第1の弁が配置されると共に、前記熱循環チャンバの流出ポートに、第2の弁が配置されており、前記第2の弁が前記第1の弁より前に閉じられることで、熱循環の前に、マイクロスフィアを、前記狭窄部から前記熱循環チャンバの本体部分に移動させる、方法。
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WO2005118867A3 (en) | 2007-12-13 |
US7799553B2 (en) | 2010-09-21 |
US8420318B2 (en) | 2013-04-16 |
KR101214270B1 (ko) | 2012-12-20 |
US20110020920A1 (en) | 2011-01-27 |
WO2005118867A2 (en) | 2005-12-15 |
US20050287572A1 (en) | 2005-12-29 |
EP1774042A2 (en) | 2007-04-18 |
KR20070030827A (ko) | 2007-03-16 |
US20120142010A1 (en) | 2012-06-07 |
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