单相裂解试剂提取总 RNA。

Isolation of Total RNA Using Monophasic Lysis Reagents.

出版信息

Cold Spring Harb Protoc. 2020 Sep 1;2020(9):101642. doi: 10.1101/pdb.top101642.

Abstract

A typical mammalian cell contains ∼10 µg of RNA, 80%-85% of which is ribosomal RNA (rRNA; chiefly the 28S, 18S, 5.8S, and 5S species). Most of the remaining 15%-20% consists of a variety of low-molecular-weight species (e.g., transfer RNAs [tRNAs] and small nuclear RNAs). These abundant RNAs are of defined size and sequence. In contrast, messenger RNA (mRNA), which makes up between 1% and 5% of the total cellular RNA, is heterogeneous in both size-from a few hundred bases to many kilobases in length-and sequence. In this introduction, the development and use of monophasic lysis reagents for the isolation of total RNA from eukaryotic cells is discussed.

摘要

典型的哺乳动物细胞含有约 10µg 的 RNA，其中 80%-85%为核糖体 RNA（rRNA；主要是 28S、18S、5.8S 和 5S 物种）。其余的 15%-20%主要由各种低分子量物种（如转移 RNA [tRNA] 和小核 RNA）组成。这些丰富的 RNA 具有明确的大小和序列。相比之下，信使 RNA（mRNA）占总细胞 RNA 的 1%至 5%，其大小和序列均具有异质性——长度从几百个碱基到几千个碱基不等。在本引言中，讨论了单相裂解试剂在从真核细胞中分离总 RNA 方面的发展和应用。