Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites

Isolation of high-quality functional RNA is prerequisite to facilitate any study related to gene expression and also for downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR), suppression subtractive hybridization (SSH) library construction, differential display (D...

Full description

Bibliographic Details
Main Author: Akshara George
Format: Article
Language:English
Published: Society for Tropical Plant Research 2018-04-01
Series:Tropical Plant Research
Subjects:
Online Access:http://www.tropicalplantresearch.com/download/247/2.pdf
id doaj-96810003d4e74075adb46128559909e8
record_format Article
spelling doaj-96810003d4e74075adb46128559909e82020-11-24T21:39:02ZengSociety for Tropical Plant ResearchTropical Plant Research2349-11832018-04-015181310.22271/tpr.2018.v5.i1.002Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolitesAkshara George0Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram-695522, Kerala, IndiaIsolation of high-quality functional RNA is prerequisite to facilitate any study related to gene expression and also for downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR), suppression subtractive hybridization (SSH) library construction, differential display (DD), real-time PCR and northern hybridization of medicinal plants. Tropical medicinal plants are rich in polysaccharides, polyphenolics and secondary metabolites that are reported to interfere with the successful RNA isolation. Conventional approaches for the extraction of functional RNA are often time-consuming and need expensive reagents. Moreover, these methods can yield only poor quality and quantity of functional RNA. In our laboratory, we have aimed at establishing a simple and efficient functional RNA extraction procedure. For this, a sodium dodecyl sulfate (SDS) based protocol free of guanidine isothiocyanate was adopted. The total extracted RNA remains in the upper aqueous phase, at acidic conditions leaving DNA and proteins in the lower organic phase. This principle is used in this protocol and the modifications made in the SDS- acid phenol RNA extraction protocol were found to be helpful for extracting RNA from tissues containing large quantities of secondary metabolites. This method can be completed within 3 hours with purity ranges from 1.8–2.0 as confirmed by A260/280 spectrophotometric readings. The above-described RNA extraction protocol works well with all the tissues examined so far, where standard RNA isolation methods failed.http://www.tropicalplantresearch.com/download/247/2.pdfFunctional RNA extractionTropical medicinal plantsPolysaccharidesPolyphenolsSecondary metabolitesRT-PCR
collection DOAJ
language English
format Article
sources DOAJ
author Akshara George
spellingShingle Akshara George
Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
Tropical Plant Research
Functional RNA extraction
Tropical medicinal plants
Polysaccharides
Polyphenols
Secondary metabolites
RT-PCR
author_facet Akshara George
author_sort Akshara George
title Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
title_short Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
title_full Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
title_fullStr Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
title_full_unstemmed Simple and efficient method for functional RNA extraction from tropical medicinal plants rich in secondary metabolites
title_sort simple and efficient method for functional rna extraction from tropical medicinal plants rich in secondary metabolites
publisher Society for Tropical Plant Research
series Tropical Plant Research
issn 2349-1183
publishDate 2018-04-01
description Isolation of high-quality functional RNA is prerequisite to facilitate any study related to gene expression and also for downstream applications such as reverse transcription-polymerase chain reaction (RT-PCR), suppression subtractive hybridization (SSH) library construction, differential display (DD), real-time PCR and northern hybridization of medicinal plants. Tropical medicinal plants are rich in polysaccharides, polyphenolics and secondary metabolites that are reported to interfere with the successful RNA isolation. Conventional approaches for the extraction of functional RNA are often time-consuming and need expensive reagents. Moreover, these methods can yield only poor quality and quantity of functional RNA. In our laboratory, we have aimed at establishing a simple and efficient functional RNA extraction procedure. For this, a sodium dodecyl sulfate (SDS) based protocol free of guanidine isothiocyanate was adopted. The total extracted RNA remains in the upper aqueous phase, at acidic conditions leaving DNA and proteins in the lower organic phase. This principle is used in this protocol and the modifications made in the SDS- acid phenol RNA extraction protocol were found to be helpful for extracting RNA from tissues containing large quantities of secondary metabolites. This method can be completed within 3 hours with purity ranges from 1.8–2.0 as confirmed by A260/280 spectrophotometric readings. The above-described RNA extraction protocol works well with all the tissues examined so far, where standard RNA isolation methods failed.
topic Functional RNA extraction
Tropical medicinal plants
Polysaccharides
Polyphenols
Secondary metabolites
RT-PCR
url http://www.tropicalplantresearch.com/download/247/2.pdf
work_keys_str_mv AT aksharageorge simpleandefficientmethodforfunctionalrnaextractionfromtropicalmedicinalplantsrichinsecondarymetabolites
_version_ 1725933073397186560