基于柱的快速方法，用于同时分离 DNA、RNA、miRNA 和蛋白质。

A column-based rapid method for the simultaneous isolation of DNA, RNA, miRNA and proteins.

机构信息

Department of Biochemistry, Banaras Hindu University, Varanasi, India.

出版信息

Cell Biol Int. 2012 Sep;36(9):779-83. doi: 10.1042/CBI20110342.

DOI:10.1042/CBI20110342

Abstract

In the 21st century, systems biology is a holistic approach to understand life by the cross-talk study between the genome, Rnome and proteome of a cell. We describe a column-based rapid method for the simultaneous extraction of DNA, RNA, miRNA (microRNA) and proteins from the same experimental sample without prior fractionation, which allows a direct correlation between genomic, epigenomic, transcriptomic and proteomic data. This method provides a simple and effective way to analyse each of these biomolecules without affecting yield and quality. We also show that isolated biomolecules are of the highest purity and compatible for all the respective downstream applications, such as PCR amplification, RT-PCR (reverse transcription-PCR), real-time PCR, reverse Northern blotting, SDS/PAGE and Western blot analysis. The buffers and reagents used in this method are optimized extensively to achieve the cost effective and reliable procedure to separate the functional biomolecules of the cell.

摘要

在 21 世纪，系统生物学是一种通过基因组、转录组和蛋白质组之间的相互作用来全面理解生命的方法。我们描述了一种基于柱的快速方法，可从同一个实验样本中同时提取 DNA、RNA、miRNA（microRNA）和蛋白质，无需事先进行分级分离，从而可以在基因组、表观基因组、转录组和蛋白质组数据之间建立直接关联。该方法为分析这些生物分子中的每一种提供了一种简单有效的方法，而不会影响产量和质量。我们还表明，分离出的生物分子具有最高的纯度，并且与所有各自的下游应用兼容，例如 PCR 扩增、RT-PCR（逆转录 PCR）、实时 PCR、反向 Northern 印迹、SDS/PAGE 和 Western blot 分析。该方法中使用的缓冲液和试剂经过了广泛的优化，以实现经济高效且可靠的程序，从而分离细胞的功能生物分子。