Simple and Efficient Room-Temperature Release of Biotinylated Nucleic Acids from Streptavidin and Its Application to Selective Molecular Detection.,Analytical Chemistry

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Simple and Efficient Room-Temperature Release of Biotinylated Nucleic Acids from Streptavidin and Its Application to Selective Molecular Detection.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-05-30 00:00:00 , DOI: 10.1021/acs.analchem.9b01873
Samuel Bearden ₁ , Fanny Wang ₁ , Adam R Hall _{1,

2}

Affiliation

The biotin–streptavidin bond is the strongest noncovalent bond in nature and is thus used extensively in biotechnology applications. However, the difficulty of releasing the bond without high temperatures or corrosive solutions can be a barrier to applications involving nucleic acids and other delicate substrates. Here, room-temperature phenol is employed to release biotin-tagged DNA constructs from streptavidin rapidly and efficiently. It is demonstrated that synthetic biotinylated DNA can be recovered at yields approaching 100% from both solution-phase and bead-bound streptavidin with as little as 12% (v/v) phenol, leaving the biotin tag active and reusable after extraction. As an application of this recovery method, biotinylated DNA fragments are isolated from a mixed solution to provide selectivity for solid-state nanopore detection.

中文翻译：

链霉亲和素在室温下简单高效地释放生物素化核酸及其在选择性分子检测中的应用。

生物素-链霉亲和素键是自然界中最强的非共价键，因此广泛用于生物技术应用。然而，在没有高温或腐蚀性溶液的情况下释放键的困难可能成为涉及核酸和其他脆弱基材的应用的障碍。此处，使用室温苯酚快速有效地从链霉亲和素中释放生物素标记的 DNA 构建体。结果表明，使用低至 12% (v/v) 的苯酚，从溶液相和珠结合的链霉亲和素中可以以接近 100% 的收率回收合成的生物素化 DNA，从而使生物素标签保持活性并在提取后可重复使用。作为该回收方法的应用，从混合溶液中分离生物素化的 DNA 片段，为固态纳米孔检测提供选择性。

更新日期：2019-05-30

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