期刊
GENETICS AND MOLECULAR RESEARCH
卷 11, 期 2, 页码 1773-1782出版社
FUNPEC-EDITORA
DOI: 10.4238/2012.June.29.10
关键词
Blackberry fruits; RNA isolation; CTAB; Acetic acid; DNA removal; Polysaccharide
资金
- National Natural Science Foundation of China [30971990]
Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was removed when acetic acid was utilized, before RNA precipitation. Thus, lithium chloride, a reagent widely used for RNA purification, was not needed. The isolation time was shortened to less than 3 h. The RNA was quite pure, with little DNA contamination. The quality of the RNA was assessed by spectrophotometric readings and electrophoresis on agarose gels. It was good enough for downstream enzymatic reactions, such as reverse transcription-PCR, cloning and real-time PCR assay. The method yielded an amount of total RNA comparable to previously described protocols.
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