JP2007057540A - 分子生物学的分析および診断用の自己アドレス可能、自己組立て小型電子システムおよびデバイス - Google Patents
分子生物学的分析および診断用の自己アドレス可能、自己組立て小型電子システムおよびデバイス Download PDFInfo
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- Mathematical Physics (AREA)
Abstract
【解決手段】 該デバイスは、微細写真平板およびマイクロ-マシーン加工の両方の技術を用いて製造することができる。特異的結合物質は、核酸およびポリペプチドのごとき分子生物学的分子を包含する。該デバイスは、分析物または反応物の輸送およびアドルスされた特異的微細-位置における反応を順次制御することができる。該デバイスは、分析物および反応物を濃縮し、非-特異的結合分子を除去し、DNAハイブリダイゼーション反応に厳格性の制御を提供し、かつ分析物の検出を改善することができる。該デバイスは、電子工学的に複製することができる。
【選択図】なし
Description
-ハイブリダイゼーションが起こった特異的微細-位置(群)から、非-特異的に結合した標的DNA配列を迅速に除去し;
-ハイブリダイゼーションが起こった特異的微細-位置(群)から、競合する相補的標的DNAを迅速に除去し;
-電位を上昇させて部分的にハイブリダイズしたDNA配列(1塩基を超える誤対合)を除去し;
-電位を調整して単一誤対合ハイブリダイゼーションに対する解析能を改善し(例えば、それにより点突然変異を同定する);
-同じバルク溶液中で起こる個々のハイブリダイゼーションに対して個々の電位を調整し;次いで -電位の調整を用いて非増幅標的DNA配列が捕捉オリゴヌクレオチド・プローブのアレイにハイブリダイズするのを改善する工程よりなる、核酸ハイブリダイゼーション反応の厳格性の制御を改善する方法をその要旨とする。
図1はマイクロリソグラフィー技術を用いて加工した自己アドレス可能な微細位置の基本設計を示す。3つの微細位置(10)(ML-1、ML-2、ML-3)が、絶縁材層/基板材上に沈着させてある金属部位(12)上の表面に形成されている。金属部位(12)は下部微細電極構造(10)として作用する。絶縁材は、金属部位(12)を相互に隔離する。絶縁材には、限定するものではないが、二酸化ケイ素、ガラス、レジスト、ゴム、プラスチックまたはセラミック材が包含される。
次いで、パッケージングしたチップは、該デバイスを制御し操作できるマイクロプロセッサー制御DC電力供給源およびマルチメーター器具に接続することができる。
マイクロリソグラフィー作製工程
一般的なマイクロリソグラフィー技術または写真平板技術を、膨大な数の小さな微細位置を有する複合「チップ」型デバイスの作製に用いることができる。デバイスの作製には複雑写真平板技術は必要でないが、材の選択、および水溶液中で電子工学的デバイスを有効に機能させるという要件には特別な配慮がまさに必要である。
この時点で、デバイス上の微細電極位置は、特殊化した浸透層および付着層で修飾する準備ができている。このことは本発明の重要な態様である。目的は、選択的な拡散特性を有する中間浸透層および最適な結合特性を有する付着表面層を微細電極上に作製することである。該付着層は特異的結合物質が付着するために、平方ミクロン(μm2)当たり105〜107個の官能基化位置を有すべきである。しかしながら、特異的結合物質の付着は、官能化から下部微細電極が保護されるよう、表面を保護膜形成または絶縁してはならない。官能化デバイスには、ある画分(〜5%ないし25%)の能動的金属微細電極表面が溶媒(H2O)分子に依然として接近でき、かつ対イオン(例えば、Na+およびCl-)および電解ガス(例えば、O2およびH2)の拡散が生じ得ることが必要である。
このセクションでは、マイクロマシーン加工技術(例えば、穿孔、粉砕等)または非リソグラフィー技術をいかに用いてデバイスを作製するかを記載する。一般的に、これらのデバイスは、マイクロリソグラフィーによって作製されるものよりも相対的に大きな微細位置(>100ミクロン)を有する。これらのデバイスは、分析適用、ならびに生体高分子合成のごとき、分取型の適用に用いることができる。大きなアドレス可能な位置は、多量の結合物質を運搬するために三次元形式(例えば、管またはシリンダー)で作製することができる。かかるデバイスは、限定するものではないが、プラスチック、ゴム、シリコン、ガラス(例えば、微細チャンネル、マイクロキャピラリー等)またはセラミックスを包含する種々の材を用いて作製することができる。マイクロマシーン加工デバイスの場合、接続回路および大きな電極構造は、当業者に知られている標準的な回路ボード・プリンティング技術を用いて材上にプリントすることができる。
-分子生物学的反応方法、例えば、制限酵素反応および分析、リガーゼ反応、キナーゼ反応、ならびに増幅方法;
-大きいまたは小さい抗原およびハプテンを含有する抗原/抗体反応;
-診断アッセイ、例えば、ハイブリダイゼーション分析、遺伝子解析、フィンガープリントおよび免疫診断;
-生体分子結合方法(すなわち、核酸、酵素、蛋白、レポーター基を有する抗体の共有結合的または非共有結合的標識);
-生体高分子合成、例えば、オリゴヌクレオチドまたはペプチドの組合せ合成;
-水溶性合成ポリマーの合成、例えば、炭化水素または直鎖状ポリアクリル酸;および -高分子分子およびナノ構造(ナノメーター・サイズの粒子および構造)の合成と作製を包含する種々の微細形式化した複数の工程および/または複合の反応ならびに方法(上記に限定されない)を迅速に行うことができる。
核酸ハイブリダイゼーションを本発明の例として用いた。なぜならば、それらは最も困難な多段階反応および多数の反応を特徴づけるからである。
(1)希釈された標的DNAおよび/またはプローブDNA配列の、ハイブリダイゼーションが起こる特異的微細位置(群)への迅速な輸送。この工程は、長くとも2〜3秒で行われる。
(2)希釈された標的DNAおよび/またはプローブDNA配列の、ハイブリダイゼーションが起こる特異的微細位置(群)での濃縮。濃縮効果は、ゆうに百万倍(>106)を超えうる。
(3)非-特異的結合標的DNA配列の、ハイブリダイゼーションが起こる特異的微細位置(群)からの迅速な除去。この工程は10ないし20秒かかる。
(4)競合する相補的標的DNA配列の、ハイブリダイゼーションが起こる特異的微細位置(群)からの迅速な除去。この工程は10ないし20秒かかる。
(6)数分のうちに完全なハイブリダイゼーション工程を行う能力。
(7)最小限の外部操作または洗浄工程でハイブリダイゼーション工程を行う能力。
(8)部分ハイブリダイズDNA配列を除去するための電子工学的な厳格性の制御(ESC)の使用。
(9)1,000〜100,000コピー範囲の非増幅ゲノム標的DNA配列のハイブリダイゼーション分析を行うことができる能力。
(10)単一塩基誤対合ハイブリダイゼーション(点突然変異)の分析を改良するためのESCの使用。
(11)マトリックス・ハイブリダイゼーションに個々の厳格性の制御を提供するためのESCの使用。
(12)非-特異的バックグラウンド成分を除去することによるハイブリダイゼーション事象の検出を改善すること。
(13)ハイブリダイゼーションを検出するために共有結合的に標識したレポータープローブまたは標的DNAを用いる必要性がない新たな方法の開発。
特異的結合物質で個々のデバイスを別々にアドレッシングすることに加えて、特異的結合物質を他のデバイスにコピーできるマスターデバイスを作製することも可能である。このことは、デバイスの作製または製造のもう1つの方法を表している。デバイス複製の工程を図13に示す。特異的結合配列でアドレスされた微細位置を含むマスターデバイスを、各々の相補的DNA配列(130)とハイブリダイズさせる。これらの相補的配列は活性化され、それ故、微細位置付着層に共有結合できる。
蛍光結合反応の場合、結合反応を分析するためにエピ蛍光型の顕微鏡検出システムを使用することができる。該システムの感度は、組み合わせたイメージング検出器エレメント(CCD、ICCD、マイクロチャネル・プレート)、またはフォトン計数PMTシステムに依存する。1の別法は、高感度CCDチップ検出器またはアバランシェ・フォトダイオード(APD)検出器をデバイスにサンドイッチ配置として直接的に組み合わせることである。さらなる別法は、デバイス中に光電子的検出器または微小電子的検出器を組み込むことである。
本発明のデバイスは、オリゴヌクレオチドおよびペプチドのごとき生体高分子の組合せ合成を行うこともできる。かかる工程によって、外部の検出または影響を全く必要としないで自己指定の合成を行うことができる。本発明の組合せ合成によって、非常に多数の配列をデバイス上で合成することができる。組合せ合成の基本概念には、輸送し、デリバリーし、濃縮し、モノマーを反応させ、試薬をカップリングし、またはアドレス可能な微細-位置で試薬を脱ブロッキングするためのデバイスの使用が含まれる。該概念はデバイスの能力を利用して、近隣の試薬および反応物の作用から特定の位置を保護する。1またはそれを超える反応物が正味正または負に荷電するか、またはこれらの工程に適したかかる試薬を作り出すかのいずれかであるこれらの化学合成プロセスにおける選択工程の同定も該概念に重要である。
合成DNAプローブを、アプライド・バイオシステムズ(Applied Biosystems)の自動DNAシンセサイザーによって通常のホスホアミダイト化学を用いて作製した。オリゴマーは5'アミノまたは3'リボヌクレオシド末端のいずれかを含むように設計された。5'の官能基を、ABI アミノリンク 2 試薬を用いることによって包含し、3'の官能基はRNA CPG 支持体から合成を開始することにより誘導した。3'リボヌクレオチド末端は過ヨウ素酸酸化法により末端のジアルデヒドに変換することが出来、これは第一アミンと反応してシッフ塩基を形成し得る。反応条件は以下の通りであった:1OD/μlの終濃度となるように水に20−30 O.D.のオリゴマーを溶解する。1容の0.1M酢酸ナトリウム、pH5.2および1容の0.45M 過ヨウ素酸ナトリウムを添加する(水中で新たに調製する)。反応を暗中室温にて少なくとも2時間撹拌しインキュベートする。反応混合物を、0.1M リン酸ナトリウム、pH7.4中で平衡化したSephadex G-10 カラム(パスツールピペット、0.6×5.5cm)の上にのせる。200μlの画分を集め、薄層クロマトグラフィー(TLC)上に2μl分をスポットし、紫外線(UV)吸収画分をためる。
以下のオリゴマーは3'リボヌクレオシド末端(U)を含む:
以下のオリゴマーが5'アミノ末端を含む:
マイクロキャピラリーチューブ(0.2mm×5mm)から微小電極を製造した。マイクロキャピラリーを0.1−1.0%のポリリジンを含む18−26%のポリアクリルアミドで満たし、多量体化させた。過剰のキャピラリーを計測し、チューブ内に気泡がトラップされるのを防ぎ、チューブ長を標準化するため除去した。キャピラリーを、共通の上部バッファー槽を共有しそれぞれ下部のバッファー槽を持つような方法で乗せた。それぞれの下部のバッファー槽は白金線電極を含んでいた。
この実施例はオリゴマーの5'末端に共有的に結合する他の付着化学を記載する。1% スクシニミジルアクリレート(分子プローブ)をポリリジンの代わりに用いる以外は上記記載のごとくキャピラリーを製造した。キャピラリーを新たに作成した、なぜならスクシミジルエステルは第一アミンと反応し、特にpH8.0以上では不安定だからである。キャピラリーを上記記載のごとく乗せ、槽を0.1M リン酸ナトリウム、pH7.4で満たした。キャピラリーを0.05mAで10分プレランする。パワーがオンの間2μl(0.1O.D.)のET10ALを上部槽にピペットで入れ、2−5分間電気泳動する。今や試験キャピラリーが負に偏るように極性を逆にしさらに2−5分間電気泳動する。結合しないDNAは反発し一方共有結合したDNAは残存する。
臭化エチジウム(EB)のごときDNA染料はDNAに挿入した場合に蛍光性となる。蛍光および結合アフィニティーはDNAが一本鎖である場合よりも二本鎖である場合の方が大きい。実施例1のごとくキャピラリーを調製し、上記記載のごとくハイブリダイズする。EBを溶液に添加し(〜0.05mMのEB終濃度)、試験キャピラリーを負に偏らせる、なぜならEBは正の電荷をおびているからである。キャピラリーを、550nmの励起および600+nmの発光におけるエピフルオレセンスによって観察した。ハイブリダイズしたおよびハイブリダイズしない微細位置が赤い蛍光を示した(EBから)。
アルミニウム(Al)および金(Au)線(0.25mm,アルドリッチ(Aldrich))をトルエン中10% 3−アミノプロピルトリエトキシシラン(APS)と反応させた。APS試薬は金属表面上で該オキシドおよび/またはヒドロキシル基と容易に反応し、オキシドおよび/またはヒドロキシル基および第一アミン基間の共有結合を形成する。アルミニウムに対する前処理は必要でなかった。
実施例5に記載されるごとくDNA−アルミニウム電極基板を調製し、ハイブリダイズさせた。ハイブリダイズした、またハイブリダイズしないDNA−Al線電極を、対照としての被覆しないAl線と共に処理した。EBを溶液に添加し、試験DNA電極を、染料を引き付けるように負に偏らせた。容器を吸引し、新しいバッファーを添加した。金属表面を顕微鏡下試験した。
該バッファーを吸引し、電極をエピフルオレセンスにより観察した。ハイブリダイズしたまたはハイブリダイズしない金属表面間の蛍光性の顕著な相違が見られるまでこれを繰り返した。
6のアドレス可能な250μmのキャピラリー配置のラジカルな列を微小−機械とした。該装置はそれぞれの微細位置におけるポテンシャルが個別にアドレス可能であるような共通の上部槽および別々の下部槽を持つ。唯一のオリゴマー結合本体が他で記載された方法によって具体的なキャピラリー微細位置に存在し、結合する。試験微細位置は正のポテンシャルを持つ一方で他方の微細位置は非特異的相互作用を防ぐため負のポテンシャルを持つ。
シリコンウエハー上の50μmの四角のアルミニウム電極の8×8マトリックス(図3参照)を設計し、製造し、スイッチボックスと包装した(詳細は装置製造の節を参照)。いくつかの物質および過程の進展が、以下に記載されるように、チップの選択性および効果を増大するためなされた。
APSの過程はチップ全体を包含する。機能化過程における選択性はチップ表面の反応性に依存した。微細位置を囲む領域の機能化および引き続くDNAの付着を減少させるため、SiO2または金属酸化物よりも、APSに対しより反応性がない物質が必要である。光レジストおよび窒化ケイ素を試した。異なるトップコートを二酸化ケイ素のチップに適用した。チップをエピフルオレセンスによって試験し、次いでAPSで処理し、続いて過ヨウ素酸酸化したポリA RNA配列(シグマ(Sigma),分子量100,000)の共有的付着が起こった。チップを、37℃にて5分間、ハイブリダイゼーションバッファー中で、テキサスレッドの200nMの溶液でラベルした20マー(T2−TR)とハイプリダイズさせた。チップをWB中で3回洗浄し、1×SSC中で1回洗浄した。チップは590nmの励起および610nmの発光での蛍光により試験した。
完成したマトリックスチップを、B&L顕微鏡およびCCDカメラに適合したプローブ・テスト・ステーション(Probe Test Station)(マイクロマニュピュレーター モデル 6000)を用いて視覚的に試験した。チップを、試験パッドと外部接触パッド間の連続性について試験した。これは、パッドを、マルチメーターに接続したマニュピュレーターのプローブチップと接触させることにより行った。連続性によって、パッドが金属表面に刻み付けられることが確実となった。パッドを次いで電気分解環境における安定性について検討した。金属線は、通常の乾燥条件で1mAまで制御されるとみなした。しかしながら、湿った環境での反応は未知であった。緩衝化された溶液(1×SSC)の一滴(1−5μl)を8×8マトリックス上にピペットで乗せた。表面張力によって、液体をその場に止め、外部接触パッド領域を乾燥した状態とする。プローブチップを接触パッドに接触させ、他のプローブチップを液体と接触させた。電流を、HP 6625A パワーサプライおよびHP3458A デジタルマルチメーターを用い、最高電圧50Vにおいて50nAまで増大させた。
実施例5に記載されるごとく、マトリックスチップをAPS試薬と共に機能化した。チップを次いで過ヨウ素酸酸化したポリA RNA(シグマ(Sigma)、平均分子量100,000)で処理した。過剰なRNAおよび結合していないRNAを除去するためチップをWB中で洗浄した。この過程により、捕捉配列を伴うチップ全体を、窒化物で被覆した領域におけるよりも暴露された金属表面において高密度で被覆した。チップを37℃にて5分間HB中T2−TRの200nMの溶液とハイブリダイズさせた。次いでWB中で3回、1×SSC中で1回、室温にてそれぞれ1分間洗浄した。チップを、590nmの励起および610nmの発光での蛍光により試験した。開かれた金属の領域は明るく蛍光性であり、パッドの形を持っていた。低い蛍光性強度および/または不規則な境界線は、パッドが完全には開いていないことを意味するであろう。付加的なプラズマエッチ時間は推奨されるであろう。
活性ハイブリダイゼーションは実施例8cからのチップを用い、1の微細位置を正に偏らせることにより行った。これは、残存する微細位置を自動的に負に偏らせるであろうスイッチボックスを用いるか、または外部溶液電極を用いることによってなされた。3マイクロリットルの水をマトリックスのパッド上に堆積させた。〜1−5nAの電流を数秒間適用し、0.1pmoleのT2−TRを溶液に添加した。液体を除去し、チップを乾燥し、テキサスレッド波長(励起590nm,発光610nm)における蛍光として試験した。正に偏った微細位置のみが蛍光性であった。これは、他の微細位置を選択的にハイブリダイズするため何回も繰り返すことができる。付加的には、1の微細位置での蛍光性DNAは、最初の配置を負に偏らせ、目的地を正に偏らせることによってもう一個の微細位置に移すことが出来る。
マトリックスを上記記載のごとくAPSで機能化した。結合本体CP−1は過ヨウ素酸酸化法によって活性化した。4の微細位置をマトリックス中正に偏らせ、残りを負に偏らせた。2マイクロリットルの水をマトリックス上に堆積させ、電流を適用した。結合本体CP−1を添加し、指示した配置において濃縮させた。液体を除去し、チップを手短かに水でリンスし、2マイクロリットルの水をチップ上に堆積させた。再び、電流を数秒間適用し、0.1pmoleのT2−TRを添加した。液体を短時間たった後に除去しチップ全体を3回WB中で洗浄した。チップを乾燥させ、蛍光を試験した。結果は、正に偏った微細位置が蛍光性であることを示す。この実施例は、特異的結合本体についての微細位置の選択的アドレッシング、配置および微細位置への配列の共有結合、被覆した微細位置への相補的標的配列の特異的なハイブリダイゼーションを示す。
3'リボヌクレオシド末端のDNA結合本体が合成され、これはHLA遺伝子dQaのポリモルフィズム(polymorphism)に特異的である。結合本体は上記記載のごとく過ヨウ素酸酸化により活性化される。逆のコンプリメントは5'アミノ末端からも合成し、他で記載したテキサスレッド、ローダミンおよびボディピイ色素のごとき発蛍光団と複合させる。微細位置は他で記載したごとくAPSでの処理によって第一アミンで機能化する。一組の1マイクロリットルの溶液をマトリックス上に置き、接触パッドは乾いたままとする。具体的な微細位置は、その微細位置を正に偏らせることによってアドレスし、過ヨウ素酸酸化したDNAオリゴマーを、〜0.1pmole添加し、その配置に移し、共有結合させた。極性を逆とし、非結合結合本体分子を除去する。これは、全ての唯一の結合本体がチップに結合するまで、他のアドレスされた微細位置の他の結合本体について繰り返される。チップを次いで、全てのアドレスされた微細位置を同時に視覚化するためと同様ひとまとめに、結合反応の特異性を決定するためのそれぞれの蛍光的にラベルしたコンプリメント配列にハイブリダイズさせる。相補的オリゴマーを除去するために変性した(0.05% SDS中90℃にて10分間)同じチップ上で、アドレスした微細位置を、ゲノムDNAのラベルしない逆のコンプリメントとハイブリダイズする。検出は、他で記載したごとく蛍光性染料検出分析によって行われる。結果は、微細位置が唯一の結合本体で特異的にアドレスされることを示すであろう。負に偏った微細位置への非特異的結合は無視できるであろう。装置および関連する結合本体化学は変性条件下で安定であり、したがって、アドレスされ製造された装置を再利用可能とする。ハイブリッドを変性する代わりの方法は、電流を増加し、および/またはそれが適用される時間を増加することであろう。
電気的強度対照に影響する装置の能力は、ラス・オンコジーン・モデル・システム(Ras oncogene model system)について示される。1の塩基対の誤対合は不都合にも融点(Tm)、すなわち二重らせんの安定性の測定値に影響する。誤対合と完全な対合(すなわち厳格性対照)を区別する伝統的な方法は、温度および塩条件に依存する。厳格性は電気泳動的ポテンシャルによっても影響され得る。以下にリストしたオリゴマーは、得られたハイブリッドが0−2の誤対合を持つように対合させることができる。オリゴマー結合本体は微細位置に結合し、他で記載したごとくハイブリダイズする。微細位置での極性は次いで逆にされ、ハイブリッドは与えられた時間定電流に付されるか、完全な対合に影響しない誤対合を変性する限定されたパワーレベルで定電流に付される。
Claims (9)
- 特異的結合DNA配列および非特異的結合DNA配列を含有する溶液からのDNAのハイブリダイゼーションを結合位置に対して電子工学的に制御する方法であって、以下の工程:
下に存在する第1の電極を包含する第1の結合位置および下に存在する第2の電極を包含する第2の結合位置に接触させて溶液を置き;
該第1の結合位置を該第2の結合位置に対して正電位に置き、該第1の位置表面上でDNAを濃縮し;次いで、
該第1の結合位置を該第2の結合位置に対して負の電位に置き、ここに、該負の電位または電流が該第1の結合位置から非特異的結合DNA配列を除去するに十分であるが特異的結合DNA配列を除去するには十分でないことを特徴とする方法。 - 特異的結合DNA配列および非特異的結合DNA配列を含有する溶液からのDNAのハイブリダイゼーションを第1および第2の結合位置に対して電子工学的に制御する方法であって、以下の工程:
第1、第2、および第3の位置に接触させて溶液を置き;
該第1および第2の結合位置を正の電位に置き、該第3の位置を負の電位に置き、該第1および第2の位置においてDNAを濃縮し;
該第1および第2の特異的結合位置を負の電位に置き、該第3の位置を正の位置に置き;次いで、
該第1および第2の結合位置を該第3の位置に対して負の電位に置き、ここに、該負の電位または電流が該第1および第2の位置から非特異的結合DNA配列を除去するに十分であるが特異的結合DNA配列を除去するには十分でないことを特徴とする方法。 - 特異的結合DNA配列および非特異的結合DNA配列を含有する溶液からのDNAのハイブリダイゼーションを第1の結合位置、次いで、第2の特異的結合位置に対して電子工学的に制御する方法であって、以下の工程:
該第1、第2、および第3の位置に接触させて溶液を置き;
該第1の結合位置を正の電位に置き、該第2の結合位置を負の電位に置き、該第1の位置においてDNAを濃縮し;
該第1の結合位置を負の電位に置き、該第2の結合位置を正の電位に置き、該第2の位置においてDNAを濃縮し;次いで、
該第1および第2の結合位置を該第3の結合位置に対して負の電位に置き、ここに、該負の電位または電流が該第1および第2の位置から非特異的結合DNAを除去するに十分であるが特異的結合DNAを除去するには十分でないことを特徴とする方法。 - 該負の電位または電流が漸次増加または減少される請求項3のハイブリダイゼーション法。
- 多数の特異的および非特異的DNA配列が結合位置のアレイに適用される請求項2または3の方法。
- DNAを溶液から複数の位置へ能動的に輸送する方法であって、以下の工程:
DNAを含有する溶液を第1、第2、第3、およびn個の位置と接触させて置き;
他の位置に対して正の電位を該第1の位置に提供し、DNAを該第1の位置に輸送し;
該第1の位置に対して正の電位を該第2の位置に提供し、DNAを該第2の位置に輸送し;
該第2の位置に対して正の電位を該第3の位置に提供し;次いで、
該プロセスをn個の位置すべてについて繰り返すことを特徴とする方法。 - 生体高分子の組み合わせ合成のための電子工学的に制御された方法であって、以下の工程:
それぞれが個々に電子工学的にアドレス可能な複数の反応位置を基板上に形成し;
各反応位置上に付着層を形成し;
該反応位置を荷電モノマー−A含有溶液と接触させて置き;
反応Aが起こるようにそれらの位置をモノマー−Aと逆の電荷に選択的に偏倚させ、反応Aが起こらないようにそれらの位置をモノマーAと同じ電荷に偏倚させ;
特定のA位置においてモノマーAを濃縮し反応させ;
未反応モノマーA含有溶液を除去し;
該反応位置を荷電モノマーB含有溶液に接触させて置き;
反応Bが起こるようにそれらの位置をモノマー−Bと逆の電荷に選択的に偏倚させ、反応Bが起こらないようにそれらの位置をモノマーBと同じ電荷に偏倚させ;
特定のB位置においてモノマーBを濃縮し反応させ;次いで、
すべての生体高分子配列が完成するまでモノマー−A、モノマー−B、ないしモノマー−Nについてn回反応を繰り返すことを特徴とする方法。 - 特定のDNA配列でアドレスされた自己アドレス可能な電子デバイスを複製する方法であって、以下の工程:
マスター自己アドレス可能電子デバイス上にアドレスされた特定のDNA配列に相補的な配列をハイブリダイゼーションさせ;
受容自己アドレス可能電子デバイス上のアドレスされていない位置を、該マスターデバイス上のアドレスされた位置に揃え;次いで、
該マスターデバイス上の位置を負に偏倚し、該受容デバイス上の位置を正に偏倚し、相補的配列を該受容デバイスに輸送させることを特徴とする方法。 - さらにマスター鋳型からの相補的配列を変性させることからなる、請求項8のパターン化配列の複製方法。
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