Title
An improved method of DNA isolation suitable for PCR-based detection of begomoviruses from jute and other mucilaginous plants
Date Issued
01 July 2009
Access level
metadata only access
Resource Type
journal article
Author(s)
Division of Crop Protection
Publisher(s)
Elsevier
Abstract
A relatively quick and inexpensive modified cetyl trimethylammonium bromide method for extraction of DNA from leaf materials containing large quantities of mucilage is described. The modification including use of more volume of extraction buffer and dissolving crude nucleic acid pellet in 1 M NaCl, reduced markedly the viscosity of the mucilage and thus in the final purification step yielded a larger quantity of mucilage-free DNA suitable for subsequent PCR-based detection of begomoviruses. The method was standardized with jute samples with yellow mosaic disease and validated with different other mucilaginous-hosts with low titre of begomoviruses. DNA isolated using this method showed consistency in yield and compatibility with PCR for detection of begomoviruses from different mucilaginous plant species. The method was compared for efficacy with other reported methods and it was found to be superior over the existing methods described for isolation of DNA from mucilaginous hosts. Thus the method described could be used on a wider scale for reliable and consistent detection of begomoviruses from mucilaginous hosts for characterization and variability study. © 2009 Elsevier B.V. All rights reserved.
Start page
34
End page
39
Volume
159
Issue
1
Language
English
OCDE Knowledge area
Protección y nutrición de las plantas
Bioquímica, Biología molecular
Subjects
Scopus EID
2-s2.0-67349224982
PubMed ID
Source
Journal of Virological Methods
ISSN of the container
01660934
Sponsor(s)
The authors are thankful to the Director, Central Research Institute for Jute and Allied Fibres for providing the infrastructural support during the present investigation. The authors are also grateful to the Indian Council of Agricultural Research, New Delhi for providing financial support in the form of project grants.
Sources of information:
Directorio de Producción Científica
Scopus