title: A modified protocol for RNA extraction from different peach tissues suitable for gene isolation and real-time PCR analysis.
abstract: RNA extraction is the first step in the study of gene isolation and expression. However, it is difficult to extract high quantity and quality RNA from tissues containing large quantities of polysaccharides and polyphenols. Peach (Prunus persica), in addition to containing high levels of polysaccharides and polyphenols, is a challenging starting material for RNA isolation using a single method because of different amounts of those substances in diverse tissues. Based on three reported methods, we developed a modified RNA isolation protocol to solve this problem, leading to high quality and quantity of total RNA from peach mesocarp tissues of fruits which were sampled from all developmental stages and different storage periods, as well as from other tissues including flowers, leaves, stems, and roots. With our modified method, 28-650 μg of total RNA was routinely obtained from per gram of fresh material, gave at least a 1.16-fold improvement by compared with those isolated by other seven methods. The RNA extracts were successfully used in downstream applications such as RT-PCR, RACE, and real-time PCR.
authors: College of Horticulture, Nanjing Agricultural University, Weigang, Nanjing 210095, Jiangsu, People's Republic of China. tongzhaoguo@163.com
authors: Zhaoguo
authors: Z
authors: Tong
authors: Shenchun
authors: S
authors: Qu
authors: Jiyu
authors: J
authors: Zhang
authors: Fei
authors: F
authors: Wang
authors: Jianmin
authors: J
authors: Tao
authors: Zhihong
authors: Z
authors: Gao
authors: Zhen
authors: Z
authors: Zhang
mesh_terms: D005638:Fruit / Q000737:chemistry / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics; D018517:Plant Roots / Q000737:chemistry / Q000235:genetics; D018547:Plant Stems / Q000737:chemistry / Q000235:genetics; D059808:Polyphenols / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis; D027861:Prunus / Q000235:genetics*; D018749:RNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D059808:Polyphenols; D011134:Polysaccharides; D018749:RNA, Plant