A simple and convenient approach for isolating RNA from highly viscous plant tissue rich in polysaccharides.
RNA isolation is a prerequisite to the study of gene expression of herbaceous plant Dendrobium nobile under an environmental stress. However, RNA isolation is difficult in the plant of genus Dendrobium, which is relatively viscous, due to being rich in polysaccharides. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to D. nobile. Here, we describe a simple and convenient method for high quality total RNA extraction from D. nobile. Main procedures are as follows: smashing plant tissue and cell wall by means of pre-cooled citrate homogenization, cleaving cell membrane by using guanidine hydrochloride lysis buffer, removing proteins, polyphenols and polysaccharides by acidic phenol/chloroform. It is particularly useful for processing large numbers of plant samples. The whole process can be completed within 2.5 h. The extracted RNA is suitable for applications such as RT-PCR, Northern analysis and screening of differential expression genes.
Li
Biao
B
Key Laboratory for Biomechanics and Tissue Engineering under the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, China. cquptlibiao@yahoo.com.cn
Wang
Bochu
B
Tang
Kun
K
Liang
Yilong
Y
Wang
Jian
J
Wei
Jinmin
J
D018929:Cell Culture Techniques; D031667:Dendrobium / Q000737:chemistry*; D018547:Plant Stems / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D012313:RNA / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014783:Viscosity
D011134:Polysaccharides; D012313:RNA