Journal
FEBS OPEN BIO
Volume 9, Issue 4, Pages 814-825Publisher
WILEY
DOI: 10.1002/2211-5463.12561
Keywords
cassava; qRT-PCR; RNA isolation; RNA sequencing
Categories
Funding
- CGIAR Research Program on Roots, Tubers and Bananas (RTB)
- CGIAR Fund Donors
- Natural Resource Institute (NRI) from the Bill & Melinda Gates Foundation (BMGF)
- MEXT KAKENHI [16H06466, 16H06464]
- JSPS
- Grants-in-Aid for Scientific Research [16H06466] Funding Source: KAKEN
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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.
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