用于DNA原位荧光标记的二部四半胱氨酸PNA探针的合成

Ge-min Fang, Oliver Seitz
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引用次数: 0

摘要

“无标记”荧光探针避免了荧光染料与寡核苷酸偶联的额外步骤或构建块,可以显着减少大量核酸样品平行生物分析的时间和成本。本文描述了一种使用固相合成和序列特异性DNA原位荧光标记的方法来合成“无标记”双钢修饰的PNA探针。这一概念是基于两个双糖修饰的肽核酸在DNA靶标上的相邻排列,形成结构优化的双糖四半胱氨酸基序,从而诱导与市购双砷染料的序列特异性荧光反应,即使在细胞裂解液等复杂介质中也是如此。该装置将帮助研究人员快速合成二部四半胱氨酸PNA探针,并开展低成本的DNA原位荧光标记实验。©2017 by John Wiley &儿子,Inc。
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Synthesis of Bipartite Tetracysteine PNA Probes for DNA In Situ Fluorescent Labeling

“Label-free” fluorescent probes that avoid additional steps or building blocks for conjugation of fluorescent dyes with oligonucleotides can significantly reduce the time and cost of parallel bioanalysis of a large number of nucleic acid samples. A method for the synthesis of “label-free” bicysteine-modified PNA probes using solid-phase synthesis and procedures for sequence-specific DNA in situ fluorescent labeling is described here. The concept is based on the adjacent alignment of two bicysteine-modified peptide nucleic acids on a DNA target to form a structurally optimized bipartite tetracysteine motif, which induces a sequence-specific fluorogenic reaction with commercially available biarsenic dyes, even in complex media such as cell lysate. This unit will help researchers to quickly synthesize bipartite tetracysteine PNA probes and carry out low-cost DNA in situ fluorescent labeling experiments. © 2017 by John Wiley & Sons, Inc.

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Current Protocols in Nucleic Acid Chemistry
Current Protocols in Nucleic Acid Chemistry Chemistry-Organic Chemistry
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期刊介绍: Published in association with International Society for Nucleosides, Nucleotides & Nucleic Acids (IS3NA) , Current Protocols in Nucleic Acid Chemistry is equally valuable for biotech, pharmaceutical, and academic labs. It is the resource for designing and running successful research projects in the rapidly growing and changing field of nucleic acid, nucleotide, and nucleoside research.
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