TITLE:
An Efficient Protocol for Total RNA Isolation from Healthy and Stressed Tissues of Mulberry (Morus sp.) and Other Species
AUTHORS:
Radha Sivarajan Sajeevan, Manchanahally Byrappa Shivanna, Karaba N. Nataraja
KEYWORDS:
Mulberry, Cardamom, Papaya, Rice, Dehydration, Salinity, RNA Isolation
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.5 No.13,
June
27,
2014
ABSTRACT:
Extraction of
quality RNA for molecular biology applications from perennial woody plants like
mulberry is complicated due to the presence of high polysaccharides,
polyphenols and other secondary metabolites. Since the existing methods failed
to yield quality RNA in sufficient quantity from leaf and root tissues of
mulberry, in this study, we modified the CTAB-based protocol. The standardised
protocol yielded high quantity (520.00 μg/g fresh weight of leaf tissue) of
quality RNA and the RNA extracted was suitable for all downstream applications such
as cDNA synthesis, PCR and whole transcriptome analysis. The method developed
was also found to be useful for isolating good quality and quantity total RNA
from desiccated and salinity stressed leaf tissues of mulberry. The protocol
was also applied successfully to isolate total RNA from leaf tissues of other
species such as cardamom, papaya and rice.