Open AccessAn optimized isolation protocol yields high‐quality RNA from cassava tissues ( Manihot esculenta Crantz)
Author(s) -
Behnam Babak,
BohorquezChaux Adriana,
CastanedaMendez Oscar Fernando,
Tsuji Hiroyuki,
Ishitani Manabu,
Becerra LopezLavalle Luis Augusto
Publication year - 2019
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12561
Subject(s) - manihot esculenta , rna , ribosomal rna , reverse transcriptase , rna extraction , 5s ribosomal rna , biology , 18s ribosomal rna , chemistry , food science , microbiology and biotechnology , biochemistry , gene , botany
We developed and modified a precise, rapid, and reproducible protocol isolating high‐quality RNA from tissues of multiple varieties of cassava plants ( Manihot esculenta Crantz). The resulting method is suitable for use in mini, midi, and maxi preparations and rapidly achieves high total RNA yields (170–600 μg·g −1 ) using low‐cost chemicals and consumables and with minimal contamination from polysaccharides, polyphenols, proteins, and other secondary metabolites. In particular, A 260 : A 280 ratios were > 2.0 for RNA from various tissues, and all of the present RNA samples yielded ribosomal integrity number values of greater than six. The resulting high purity and quality of isolated RNA will facilitate downstream applications (quantitative reverse transcriptase‐polymerase chain reaction or RNA sequencing) in cassava molecular breeding.