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Maxam-Gilbert Sequencing01:05

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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Next-generation Sequencing03:00

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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Sanger Sequencing01:57

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DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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核酸反応経路の制限されたマルチステートシーケンスの設計

Brian R Wolfe1, Nicholas J Porubsky2, Joseph N Zadeh1

  • 1Division of Biology & Biological Engineering, California Institute of Technology , Pasadena, California 91125, United States.

Journal of the American Chemical Society
|February 14, 2017
PubMed
まとめ
この要約は機械生成です。

この研究は,核酸配列を設計するための計算フレームワークを導入し,それらの混合反応を制御します. この方法は,特定の反応経路のための配列を最適化し,正確な分子プログラミングと合成生物学アプリケーションを可能にします.

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科学分野:

  • バイオテクノロジー
  • コンピュータ生物学
  • 合成生物学

背景:

  • 複雑な相互作用のための核酸配列の設計は困難です.
  • 特定の反応経路を制御するには 精密な配列工学が必要です

研究 の 目的:

  • 制御された混合化と反応経路のための核酸配列の設計のための計算的枠組みを提示する.
  • 分子プログラミングと合成生物学システムの 精密な工学を可能にします

主な方法:

  • マルチステート最適化問題としてシーケンスの設計を策定する.
  • 反応物,中間物,および産物状態を表す標的試験管を使用する.
  • 経路内の反応と経路外の反応を制御するために,ポジティブとネガティブの両方の設計パラダイムを組み込む.
  • 構成,補完性,パターン防止,生物学的制約を含むユーザー指定の制約を適用します.

主要な成果:

  • 処方された反応経路で核酸配列を設計するための枠組みを開発した.
  • 特定の二次構造と濃度のために設計する能力を示した.
  • 望ましくないオフターゲットの相互作用に対して明示的な設計を有効にしました.
  • 複雑なアプリケーションのための制限されたマルチステートシーケンスの設計を容易にした.

結論:

  • 制限された多状態配列設計フレームワークは,核酸反応の精密な工学を可能にします.
  • このアプローチは分子プログラミングと合成生物学における 多様な応用をサポートしています
  • NUPACKのウェブアプリケーションは,この設計ツールへのオンラインアクセスを提供しています.