32P- and biotin-labelled in vitro transcribed cRNA probes for the detection of potato spindle tuber viroid and chrysanthemum stunt viroid.
Replacing nick-translated DNA probes by in vitro transcribed complementary RNA (cRNA) probes considerably increased the sensitivity of dot-blot detection tests of potato spindle tuber viroid and chrysanthemum stunt viroid. As compared to the limit of detection of 5-10 pg of viroid obtained with 32P-labelled DNA probes, cRNA probes allow the detection of less than 1 pg of pure viroid. When labelled with biotin by incorporation of biotin-labelled ribonucleotides, the cRNA probes have a limit of detection of approximately 5 pg of purified viroid.
Candresse
T
T
INRA, Centre de Recherches de Bordeaux, Pont de la Maye, France.
Macquaire
G
G
Brault
V
V
Monsion
M
M
Dunez
J
J
D001710:Biotin; D015336:Molecular Probe Techniques; D010761:Phosphorus Radioisotopes; D010935:Plant Diseases; D012313:RNA / Q000235:genetics; D015347:RNA Probes; D018075:RNA, Complementary; D012367:RNA, Viral / Q000235:genetics; D014772:Viroids / Q000235:genetics / Q000302:isolation & purification*
D010761:Phosphorus Radioisotopes; D015347:RNA Probes; D018075:RNA, Complementary; D012367:RNA, Viral; D012313:RNA; D001710:Biotin