An improved CTAB-ammonium acetate method for total RNA isolation from cotton

Phytochemical Analysis : PCA
Lu ZhaoXin-Qiang He

Abstract

Cotton is an important economic crop. Genetic, developmental and molecular studies of cotton require high-quality total RNA from different tissues. Due to the richness in polyphenols and polysaccharides, the Trizol-based methods and other commercial kits are unsuitable for RNA isolation from cotton. Available methods are generally laborious and time-consuming. To develop an easy, simple and rapid cetyltrimethylammonium bromide (CTAB)-ammonium acetate protocol that takes less time and obtains high yield and quality of RNA from polysaccharide- and polyphenol-rich cotton tissues. Based on the original CTAB protocol, we used phenol-chloroform and chloroform-isoamyl alcohol to remove proteins, polysaccharides and polyphenols, and ammonium acetate to precipitate RNA, reducing the incubation time prior to RNA precipitation. After adding ammonium acetate to precipitate RNA, all centrifugation steps (14000 × g) were carried out at 4°C to avoid degradation. The procedure took only 1.5 h and was suitable for different cotton tissues. The A(260) : A(280) ratios ranged from 1.80 to 1.85 with clear 28 s and 18 s ribosomal RNA bands in 1.2% agarose gel. The isolated RNA was usable for downstream molecular studies, such as reverse transcriptio...Continue Reading

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Citations

Jan 10, 2014·Preparative Biochemistry & Biotechnology·Rafael da Silva MessiasCesar Valmor Rombaldi
Feb 26, 2016·Plant Biotechnology Journal·Xiu-Ming WuXiao-Ya Chen
Oct 13, 2018·Thoracic Cancer·Miao Chen, Songbo Xie
Sep 10, 2014·Proteomics. Clinical Applications·Sarah PlumCaroline May
Feb 21, 2019·Journal of Cellular Biochemistry·Jingrui Li, Min Liu
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Feb 23, 2020·Frontiers in Plant Science·Feifei Qi, Fuxin Zhang
Jul 27, 2021·PloS One·Paola de Avelar CarpinettiMarcia Flores da Silva Ferreira

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