A green method of staining DNA in polyacrylamide gel electrophoresis based on fluorescent copper nanoclusters synthesized in situ

A green method of staining DNA in polyacrylamide gel electrophoresis based on fluorescent copper nanoclusters synthesized in situ
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基于原位合成荧光铜纳米簇的聚丙烯酰胺凝胶电泳DNA染色绿色方法

DOI:
10.1007/s12274-015-0778-y
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发表时间:
2015-08
期刊:
影响因子:
9.9
通讯作者:
Li Genxi
Li Genxi
中科院分区:
材料科学1区
文献类型:
--
作者:
Zhu Xiaoli;Shi Hai;Shen Yalan;Zhang Bin;Zhao Jing;Li Genxi

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核酸染色染料的安全性一直被认为是一个问题。为寻找最流行但有毒的染色染料溴化乙啶(EtBr)的替代品,人们作出了广泛的努力。然而,到目前为止,还没有一种能够保证足够安全的染色方法。本文报道了一种在聚丙烯酰胺凝胶电泳中对DNA进行绿色染色的方法,该方法在凝胶中原位合成了DNA模板化的荧光铜纳米簇(CuNCs),使DNA条带在紫外线下可见。此外,对该染色方法的性能进行了全面的研究,实验结果表明该染色方法具有良好的灵敏度、稳定性和可用性。同时,在我们的动物实验中,两种试剂(硫酸铜和抗坏血酸)以及合成的CuNCs已被证明与皮肤接触无毒。此外,这项工作中使用的所有试剂都很容易获得,成本低,操作简单。因此,这种基于原位合成dna模板化荧光cccs的新型染色方法具有许多潜在的应用前景。
The safety of nucleic acid staining dyes has long been recognized to be a problem. Extensive efforts have been made to search for alternatives to the most popular but toxic staining dye, ethidium bromide (EtBr). However, so far no staining method that can be guaranteed to be sufficiently safe has been developed. In this paper, we report a green staining method of DNA in polyacrylamide gel electrophoresis, where in situ synthesis of DNA-templated fluorescent copper nanoclusters (CuNCs) in the gel is achieved to make the DNA bands visible under UV light. Moreover, a comprehensive study of the performance of this staining method has been conducted and the experimental results show that it has favorable sensitivity, stability, and usability. Meanwhile, in our animal experiments, the two reagents (copper sulfate and ascorbic acid) as well as the synthesized CuNCs have been proven to be non-toxic in contact with skin. In addition, all the reagents employed in this work are readily available and low cost, and the procedure is simple to carry out. Therefore, this novel staining method based on the in situ synthesis DNA-templated fluorescent CuNCs has many potential applications.
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