PMID: 3755443May 1, 1986Paper

Detection of citrus exocortis viroid in crude extracts by dot-blot hybridization: conditions for reducing spurious hybridization results and for enhancing the sensitivity of the technique

Journal of Virological Methods
R Flores

Abstract

Dot-blot assays to detect citrus exocortis viroid (CEV), in clarified sap and unfractionated total nucleic acid preparations of CEV-infected Gynura aurantiaca and chrysanthemum, were impaired by the non-specific binding of the radioactive probe shown by the healthy controls. This non-specific background was considerably reduced by the addition to the hybridization mixture, of the fraction of nucleic acids from healthy plants which are insoluble in 2 M LiCl (containing mainly the large ribosomal RNAs). Sample denaturation with formaldehyde was found to provide a high increase of hybridization, when compared with samples either denatured with formamide or directly spotted. Nitrocellulose was observed to be a better solid support than charge-modified nylon, in terms of the sensitivity of viroid detection by spot hybridization.

References

Sep 6, 1976·Biochimica Et Biophysica Acta·J M TaylorJ Summers
Jan 1, 1985·Journal of Virological Methods·M Bar-JosephA Rosner
Jun 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·R M Franklin
Oct 15, 1983·Analytical Biochemistry·A GranellV Conejero
May 1, 1984·Analytical Biochemistry·J Meinkoth, G Wahl
Jul 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·J S Semancik, K L Harper

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Citations

Apr 1, 1989·Journal of Virological Methods·M WełnickiW Zagórski
Jan 1, 1990·Research in Virology·R FloresG Llácer
Aug 15, 1998·Journal of Virological Methods·B NavarroR Flores
Dec 20, 1994·Proceedings of the National Academy of Sciences of the United States of America·J A DaròsR Flores
Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·J A Daròs, R Flores
Aug 18, 1999·Proceedings of the National Academy of Sciences of the United States of America·M de la PeñaR Flores
Feb 13, 2008·European Archives of Oto-rhino-laryngology : Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery·Petros V VlastarakosThomas P Nikolopoulos
Nov 24, 2007·European Archives of Oto-rhino-laryngology : Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery·Petros V VlastarakosGeorge Kreatsas
Jan 1, 1994·Psychophysiology·G G BerntsonV T Fabro

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