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从 TRIzol 中提取 RNA 和 DNA 后,可高效回收骨骼肌蛋白质。

High-yield skeletal muscle protein recovery from TRIzol after RNA and DNA extraction.

机构信息

Department of Physiology, College of Medicine, University of Kentucky, Lexington, KY, USA.

Center for Muscle Biology, University of Kentucky, Lexington, KY, USA.

出版信息

Biotechniques. 2020 Oct;69(4):264-269. doi: 10.2144/btn-2020-0083. Epub 2020 Aug 11.

Abstract

Extraction of DNA, RNA and protein from the same sample would allow for direct comparison of genomic, transcriptomic and proteomic information. Commercially available kits exhibit poor protein yield and the TRIzol reagent produces a protein pellet that is extremely difficult to solubilize. In response to these limitations, this study presents an optimized method for the extraction of protein from the organic phase of TRIzol that allows for higher yield recovery of skeletal muscle protein compared with direct homogenization in a common protein lysis buffer. The presented method is inexpensive, simple and fast, requires no additional treatment of the protein pellet for dissolution, and is compatible with downstream western blot applications.

摘要

从同一个样本中提取 DNA、RNA 和蛋白质,可以直接比较基因组、转录组和蛋白质组信息。市售试剂盒的蛋白质产率较差,而 TRIzol 试剂产生的蛋白质沉淀极难溶解。针对这些限制,本研究提出了一种从 TRIzol 的有机相中提取蛋白质的优化方法,与直接在常用的蛋白质裂解缓冲液中匀浆相比,该方法可提高骨骼肌蛋白的产量回收率。所提出的方法经济、简单、快速,无需对蛋白质沉淀进行额外的溶解处理,并且与下游的 Western blot 应用兼容。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb7d/7566772/45af57d49a60/btn-69-264-g1.jpg

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