title: Isolation of High-Quality Total RNA from Chinese Fir (Cunninghamia lanceolata (Lamb.) Hook).
abstract: RNA isolation with RNA in a high quantity is a basic analytical method in plant genetics, molecular biology and related physiological investigations. To understand the genetic and molecular biology of Chinese fir, sufficient high-quality total RNA must be obtained for cDNA library construction and other downstream molecular applications. However, extracting RNA from Chinese fir is difficult and often requires the modification of existing protocols. Chinese fir tissues containing large amounts of polysaccharides and polyphenol compounds and are one of the most difficult plant tissues for RNA isolation. Therefore, we developed a simple method for extracting high-quality RNA from Chinese fir tissues. RNA isolations were performed within two hours, RNA quality was measured for yield and purity. Total RNA obtained from this procedure was successfully used for cDNA library construction, RT-PCR and transcriptome sequencing. It was proven that extracted RNA was intact and suitable for downstream molecular applications, including RT-PCR and qPCR, and other downstream molecular applications. Thus, this protocol represents a simple, efficient, and low-cost method.
authors.affiliation: College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China; State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Ma
authors.affiliation: State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China; College of Forestry, Fujian Agricultural and Forestry University, Fuzhou, China.
authors.forename: Binlong
authors.initials: B
authors.lastname: Huang
authors.affiliation: State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China; College of Forestry, Fujian Agricultural and Forestry University, Fuzhou, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Xu
authors.affiliation: State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China; College of Forestry, Fujian Agricultural and Forestry University, Fuzhou, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China; College of Forestry, Fujian Agricultural and Forestry University, Fuzhou, China.
authors.forename: Shubin
authors.initials: S
authors.lastname: Li
authors.affiliation: State Forestry Administration Engineering Research Center of Chinese Fir, Fuzhou, China; College of Forestry, Fujian Agricultural and Forestry University, Fuzhou, China.
authors.forename: Sizu
authors.initials: S
authors.lastname: Lin
mesh_terms: D005591:Chemical Fractionation / Q000379:methods*; D032486:Cunninghamia / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D011786:Quality Control; D018749:RNA, Plant / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D018749:RNA, Plant