An optimized isolation protocol yields high-quality RNA from cassava tissues (Manihot esculenta Crantz)

被引:16
|
作者
Behnam, Babak [1 ]
Bohorquez-Chaux, Adriana [2 ]
Fernando Castaneda-Mendez, Oscar [2 ]
Tsuji, Hiroyuki [1 ]
Ishitani, Manabu [2 ]
Becerra Lopez-Lavalle, Luis Augusto [2 ]
机构
[1] Yokohama City Univ, Kihara Inst Biol Res, Yokohama, Kanagawa, Japan
[2] Int Ctr Trop Agr CIAT, Recta Cali Palmira Km 17, Valle Del Cauca, Colombia
来源
FEBS OPEN BIO | 2019年 / 9卷 / 04期
基金
比尔及梅琳达.盖茨基金会;
关键词
cassava; qRT-PCR; RNA isolation; RNA sequencing; EFFICIENT PROTOCOL; TRANSCRIPTOME ANALYSIS; FUNCTIONAL RNA; EXTRACTION; DROUGHT; RICH;
D O I
10.1002/2211-5463.12561
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (170-600gg(-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.
引用
收藏
页码:814 / 825
页数:12
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