Open AccessModified CTAB and TRIzol protocols improve RNA extraction from chemically complex Embryophyta
Author(s) -
JordonThaden Ingrid E.,
Chanderbali Andre S.,
Gitzendanner Matthew A.,
Soltis Douglas E.
Publication year - 2015
Publication title -
applications in plant sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 23
ISSN - 2168-0450
DOI - 10.3732/apps.1400105
Subject(s) - trizol , biology , rna , rna extraction , dna , transcriptome , chromatography , extraction (chemistry) , computational biology , gene , biochemistry , gene expression , chemistry
Premise of the study: Here we present a series of protocols for RNA extraction across a diverse array of plants; we focus on woody, aromatic, aquatic, and other chemically complex taxa. Methods and Results: Ninety‐one taxa were subjected to RNA extraction with three methods presented here: (1) TRIzol/TURBO DNA‐ free kits using the manufacturer's protocol with the addition of sarkosyl; (2) a combination method using cetyltrimethylammonium bromide (CTAB) and TRIzol/sarkosyl/TURBO DNA‐ free ; and (3) a combination of CTAB and QIAGEN RNeasy Plant Mini Kit. Bench‐ready protocols are given. Conclusions: After an iterative process of working with chemically complex taxa, we conclude that the use of TRIzol supplemented with sarkosyl and the TURBO DNA‐ free kit is an effective, efficient, and robust method for obtaining RNA from 100 mg of leaf tissue of land plant species (Embryophyta) examined. Our protocols can be used to provide RNA of suitable stability, quantity, and quality for transcriptome sequencing.