Using silica particles to isolate total RNA from plant tissues recalcitrant to extraction in guanidine thiocyanate.
The most commonly used protocol of the RNA isolation, the guanidine thiocyanate method, was unsuitable for recalcitrant plant tissues containing a large amount of storage proteins and secondary metabolites. We demonstrated that RNA could bind to the silica particles, which have been used successfully in DNA isolation from various sources, under a high concentration of NaCl in the presence of ethanol and sodium acetate. Based on this observation, an efficient, inexpensive, and highly reproducible technique, the acid phenol-silica method, was developed to isolate high-quality RNAs from various plant tissues recalcitrant to extraction in guanidine thiocyanate.
Ding
Ling-Wen
LW
Key Laboratory of Gene Engineering of the Ministry of Education and State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Sun
Qiao-Yang
QY
Wang
Zhao-Yu
ZY
Sun
Yong-Bin
YB
Xu
Zeng-Fu
ZF
D004587:Electrophoresis, Agar Gel; D006146:Guanidines / Q000737:chemistry*; D010936:Plant Extracts / Q000737:chemistry*; D010944:Plants / Q000737:chemistry* / Q000235:genetics; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*; D013861:Thiocyanates / Q000737:chemistry*
D006146:Guanidines; D010936:Plant Extracts; D013861:Thiocyanates; C054436:guanidine thiocyanate; D012313:RNA; D012822:Silicon Dioxide