A simple phenol-free isolation method for high-quality RNA from bilberry

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Wang , K , Liu , M , Cui , F & Asiegbu , F 2021 , ' A simple phenol-free isolation method for high-quality RNA from bilberry ' , MethodsX , vol. 8 , 101481 . https://doi.org/10.1016/j.mex.2021.101481

Title: A simple phenol-free isolation method for high-quality RNA from bilberry
Author: Wang, Kai; Liu, Mengxia; Cui, Fuqiang; Asiegbu, Fred
Contributor organization: Department of Forest Sciences
Helsinki Institute of Sustainability Science (HELSUS)
Viikki Plant Science Centre (ViPS)
Frederick Asiegbu / Principal Investigator
Forest Ecology and Management
Date: 2021-08-09
Language: eng
Number of pages: 5
Belongs to series: MethodsX
ISSN: 2215-0161
DOI: https://doi.org/10.1016/j.mex.2021.101481
URI: http://hdl.handle.net/10138/334337
Abstract: High-quality RNA is required for accurate gene expression and transcriptome analysis. The current methods of RNA extraction from berry fruits are either time-consuming or expensive. To simplify the conventional phenol chloroform based RNA extraction method, we modified the protocol with less steps as well as the removal of the use of phenol. In this protocol, the extraction buffer is composed of hexadecyltrimethyl ammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and Dithiothreitol (DTT). The method facilitates efficient removal of polysaccharides and phenolic compounds from both fruit pulp and fruit peel. Additionally, the protocol is phenol free and less toxic than traditional phenol-containing method. High-quality RNA, with RNA Integrity Number value > 8, isolated by this method is applicable for RNA sequencing and qPCR. Only 3-4 working hours are required for one batch of RNA isolation. Our method replaces the use of phenol-chloroform with chloroform, making the extraction less toxic. The bilberry friut RNA is of high-quality and purity with less time input. (C) 2021 The Authors. Published by Elsevier B.V.
Subject: 11831 Plant biology
RNA isolation
Fruit pulp
Fruit peel
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion

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