A fast, sensitive and specific method for rice dwarf virus detection by northern blot hybridization.
Based on stability of double-stranded (ds) RNA, a new, fast, sensitive, and specific method for detection of genomic rice dwarf virus (RDV) dsRNA by molecular hybridization was developed. In contrary to the commonly used, standard Northern blot analysis, dsRNA is denatured in the immobilized state on the blot. Therefore, risk of degradation of single-stranded (ss) RNA by ribonuclease (RNase) during sample preparation, electrophoresis and blotting is eliminated. This method overcomes disadvantage of incomplete denaturation of dsRNA in Northern blot analysis. In conclusion, the newly developed method is reliable, sensitive, very specific, and gives a low background. The entire procedure is also less time consuming; it can be completed within 2-3 days. The new method may be regarded as a modification of the standard Northern blot analysis.
Yu
L
L
College of Life Science, Peking University, Beijing, P.R. of China. liyi@pku.edu.cn
Li
Y
Y
Gu
G
G
Wang
Y
Y
Zhou
B
B
An
C
C
Chen
Z
Z
D015152:Blotting, Northern; D009693:Nucleic Acid Hybridization; D012275:Oryza / Q000821:virology*; D012330:RNA, Double-Stranded / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis*; D012087:Reoviridae / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods
D012330:RNA, Double-Stranded; D012367:RNA, Viral