Rapid isolation of high-quality total RNA from taxus and ginkgo.
An easy and efficient protocol was developed for isolating good-quality total RNA from various tissues including fruits, leaves, stems, and roots of ancient gymnosperm species, taxus and ginkgo. The protocol was developed based on the CTAB method with modifications, including higher-strength CTAB to help the lysis of plant cells, more PVP, and beta-mercaptoethanol to prevent oxidation of phenolic complexes, and higher-centrifugation force to get rid of most cell debris and to ensure RNA quality. In RNA isolation, chloroform/isoamyl alcohol was used to remove proteins, genomic DNA, and secondary metabolites and lithium chloride was subsequently adopted to concentrate total RNA away from most of the cytoplasmic components. Good-quality total RNA from various tissues of native taxus and ginkgo could be easily isolated within 24 hr by this protocol which avoided the limitation of plant materials and the usage of dangerous chemicals, such as phenol, and could provide total RNA for all kinds of further molecular studies.
State Key Laboratory of Genetic Engineering, School of Life Sciences, Morgan-Tan International Center for Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Fudan University, Shanghai, P.R. China.
Zhihua
Z
Liao
Min
M
Chen
Liang
L
Guo
Yifu
Y
Gong
Feng
F
Tang
Xiaofen
X
Sun
Kexuan
K
Tang
D020441:Ginkgo biloba / Q000737:chemistry*; D018514:Plant Structures / Q000737:chemistry*; D012313:RNA / Q000302:isolation & purification*; D020946:Taxus / Q000737:chemistry*
D012313:RNA