JP2016511001A - 核酸配列決定機器中のフローセルを浄化するための装置及び方法 - Google Patents
核酸配列決定機器中のフローセルを浄化するための装置及び方法 Download PDFInfo
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Abstract
Description
本出願は、2014年3月14日に提出された、表題「核酸配列決定装置のフローセルを浄化するための装置及び方法」の米国出願番号13/802,957の利益に関連し、主張するものであり、その内容は、完全に及び全ての目的のために参照によって本明細書に取り入れられる。
過去25年に渡って、産生されてジーンバンク(GeneBank)に蓄積されたDNA配列情報の量は、指数関数的に成長してきた。多くの次世代配列決定技術は、合成による配列決定(SBS)の形態を使用し、特に、制御された方法において、チップに結合された単鎖DNAの鋳型の配列を読み取るために、デザインされたヌクレオチド及びDNAポリメラーゼが使用される。他の次世代配列決定技術は、核酸配列を決定するために、天然ヌクレオチド及び/又はポリメラーゼ、あるいは、標識されたオリゴヌクレオチド及びライゲーション酵素が使用されうる。高スループットを達成するために、それぞれが単独又は複数の分子である、何百万ものこのような鋳型スポットが、配列決定チップ全体に配列され、それらの配列は独立して読み取られ、記録される。高スループット配列決定を実施するという欲求は、より速く処理し、コストを下げるという要求から生じる。しかしながら、商業的な高スループットシステムは、大規模な配列決定(例えば、10〜100ギガ塩基)のコストを下げる一方、より小規模な配列決定(例えば、100メガ塩基〜1ギガ塩基)では、コストがかかり、不都合なものにする。
一実施形態において、1又は数個の可動式フローセルを有する核酸配列決定を行う装置を操作するための方法が提供される。該方法は、入口ポート及び出口ポートを備えた流路を有するフローセル提供すること、及び、第一液体試薬で該入口ポートから該出口ポートまでの流路を満たすこと、を含む。該方法は、入口通路及び出口通路を備えたステーションブロックを提供すること、並びに、入口通路と流体連通している入口ポートと出口通路と流体連通している出口ポートに配置するために該ステーションブロック上にフローセルを置くことも含んでいる。該方法は、ガス状の泡を入口ポートへ導入し、該ガス状の泡を、流路を通って、出口通路へと移動するため、及び、第二液体試薬で流路を満たすために、入口通路から入口ポートへ第二液体試薬を運搬すること、及び、ステーションブロックからフローセルを取り除くことも含んでいる。
本明細書中で述べる代表的な実施態様は、小規模の合成による配列決定(SBS)装置で使用されるフローセルから、試薬を浄化するための装置及び方法に関する。本明細書中で記述される実施態様は、前述の出願において特に有用であると考えられる一方、該装置及び方法の他の使用は、本開示の観点及び本発明の実施において、当業者にとっては明らかでありうる。例えば、実施態様は、流体がフローセルに導入される処理するステーションブロックから定期的にフローセルを解放する他の種類の装置において有用であることを見出しうる。このように、以下の例は、本発明の範囲の例を限定することを意図するものではない。
Claims (21)
- 入口ポート及び出口ポートを備えた流路を含むフローセルを提供すること;
該入口ポートから該出口ポートまで、第一液体試薬で、該流路を充填すること;
入口通路及び出口通路を備えたステーションブロックを提供すること;
該入口通路と流体連通した該入口ポート及び該出口通路と流体連通した該出口ポートを配置するために、該ステーションブロック上に該フローセルを載せること;
気泡を該入口ポートへ導入すること;
該気泡が該流路を通って該出口通路へと移動するため、及び、第二液体試薬で該通路を充填するために、該入口通路から該入口ポートへ第二液体試薬を移動すること;及び、
該ステーションブロックから該フローセルを除くこと、
を含む、1又は数個の可動式フローセルを備えた核酸配列決定装置を操作する方法。 - 該気泡を該入口ポートへと導入すること、が:
該ステーションブロック上に該フローセルを載せる前に、該入口通路の上端において、気体スペースを提供すること;及び、
それによって気泡を形成するために、該フローセルを該ステーションブロックに載せる際に、該気体スペース中の気体を捕捉すること、
を含む、請求項1に記載の方法。 - 該気泡を該入口ポートへと導入することが、該第二試薬の前の該入口ポートへ該気泡を移動するために、該第二液体試薬を該入口ポートへ移動すること、をさらに含む、請求項2に記載の方法。
- 該入口通路の該先端における該気体スペースを提供することが、該ステーションブロック上に該フローセルを載せる前に、該第二液体試薬の一部を蒸発すること、を含む、請求項2に記載の方法。
- 該第二液体試薬の該一部を蒸発することが、約7.5〜約15秒の期間、華氏約140度〜150度の温度で、該ステーションブロックを加熱すること、を含む、請求項4に記載の方法。
- 該入口通路の該先端に該気体スペースを提供することが、該ステーションブロック上に該フローセルを載せる前に、加圧した気体を該入口通路へ運搬すること、を含む、請求項2に記載の方法。
- 該入口通路の該先端に該気体スペースを提供することが、該ステーションブロック上に該フローセルを載せる前に、該入口通路の該先端から該第二液体試薬の一部を排出すること、を含む、請求項2に記載の方法。
- 該入口通路の該先端に該気体スペースを提供することが、該ステーションブロック上に該フローセルを載せる前に、該入口通路の該先端から該第二液体試薬の一部を汲み上げること、を含む、請求項2に記載の方法。
- 該入口通路の該先端から該第二液体試薬の一部を汲み上げることが、可変−置換チャンバを用いて該第二液体試薬の該一部を除去すること、を含む、請求項8に記載の方法。
- 該気泡が該流路を通って該出口通路へと移動するため、及び、該第二液体試薬で該流路を充填するために、該入口通路から該入口ポートへ該第二液体試薬を移動することが、該出口通路の下流に配置された吸い上げポンプを操作すること、を含む、請求項1に記載の方法。
- 入口ポート及び出口ポートを備えた流路を含むフローセルを提供すること;
第一入口通路及び第一出口通路を備えた第一ステーションブロックを提供すること;
該第一入口通路と流体連通した該入口ポート及び該第一出口通路と流体連通した該出口ポートを配置するために、該第一ステーションブロック上に該フローセルを載せること;
該流路を該第一液体試薬で満たすために、該第一入口通路から該入口ポートへ、第一液体試薬を汲み上げること;
該第一ステーションブロックから該第一フローセルを除去すること;
入口ポート及び出口ポートを備えた第二ステーションブロックを提供すること;
該第二入口通路と流体連通した該入口ポート及び該第二出口通路と流体連通した該出口ポートを配置するために、該第二ステーションブロック上に該フローセルを載せること;
気泡を該入口ポートへ導入すること;
該気泡が該入口ポートから該第二出口通路へ移動するため、及び、第二液体試薬で該流路を充填するために、該第二入口通路から該入口ポートへ第二液体試薬を汲み上げること;及び、
該ステーションブロックから該フローセルを除くこと、
を含む、1又は数個の可動式フローセルを備えた核酸配列決定装置を操作する方法。 - 該気泡を該入口ポートへ導入することが;
該第二ステーションブロック上に該フローセルを載せる前に、該第二入口通路の上端で気体スペースを提供すること;及び
該気泡を形成するために、該第二ステーションブロックへ該フローセルを載せる際に、該気体スペース中のガスを捕捉すること、を含む請求項11に記載の方法。 - 該気泡を該入口ポートへ導入することが、該第二液体試薬の前の該入口ポートへ該気泡を移動するために、該第二液体試薬を該入口ポートへ汲み上げること、をさらに含む、請求項12に記載の方法。
- 該入口通路の該先端の気体スペースを提供することが、該第二ステーションブロック上に該フローセルを載せる前に、該第二液体試薬の一部を蒸発すること、を含む、請求項12に記載の方法。
- 該第二液体試薬の一部を蒸発することが、約7.5〜約15秒の期間、華氏約140度〜150度の温度で、該ステーションブロックを加熱すること、を含む、請求項14に記載の方法。
- 該第二入口通路の該先端に該気体スペースを提供することが、該第二ステーションブロック上に該フローセルを載せる前に、加圧した気体を該入口通路へ運搬すること、を含む、請求項12に記載の方法。
- 該第二入口通路の該先端に該気体スペースを提供することが、該第二ステーションブロック上に該フローセルを載せる前に、該第二入口通路の該先端から該第二液体試薬の一部を吸い取ること、を含む、請求項12に記載の方法。
- 該第二入口通路の該先端に該気体スペースを提供することが、該第二ステーションブロック上に該フローセルを載せる前に、該第二入口通路の該先端から該第二液体試薬の一部を汲み上げること、を含む、請求項12に記載の方法。
- 該第二入口通路の該先端から該第二液体試薬の一部を汲み上げることが、可変−置換チャンバを用いて該第二液体試薬の一部を除去すること、を含む、請求項18に記載の方法。
- 該気泡が該流路を通って出口通路へ移動するため、及び、第二液体試薬で該流路を充填するために、該第二入口通路から該入口ポートへ第二液体試薬を汲み上げることが、該出口通路の下流に配置された汲み上げポンプを操作すること、を含む、請求項11に記載の方法。
- 第一液体試薬を汲み上げる工程、及び、第二液体試薬を汲み上げる工程が、第一出口通路及び第二出口通路に流体連通された一つの汲み上げポンプを操作すること、を含む、請求項11に記載の方法。
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US20140259607A1 (en) | 2014-09-18 |
EP2971176A4 (en) | 2016-11-16 |
WO2014159409A1 (en) | 2014-10-02 |
US9146248B2 (en) | 2015-09-29 |
CN105492626A (zh) | 2016-04-13 |
CN105492626B (zh) | 2018-05-04 |
EP2971176A1 (en) | 2016-01-20 |
AU2014240564A1 (en) | 2015-10-15 |
AU2014240564B2 (en) | 2019-11-21 |
JP6495885B2 (ja) | 2019-04-03 |
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