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One economical extraction method to detect all graft transmissible pathogens of citrus

One economical extraction method to detect all graft transmissible pathogens of citrus

Elharony, S.B.1,3, SaharA.Youssef2 and Richard F. Lee3

1Citrus Department, Horticulture Research Institute, Agriculture Research Center, Giza,
Egypt, 2 Virus and Phytoplasma Department, Plant Pathology Research Institute, Agriculture
Research Center, Giza, Egypt, 3United State Department of Agriculture-Agriculture
Research Service National Clonal Germplasm Respiratory of Citrus and Dates, Riverside,
California, United State of America

ABSTRACT

A simple and applicable extraction method for the isolation of
high quality nucleic acid (DNA and RNA) to detect all graft
transmissible pathogens of citrus including prokaryotic pathogens,
DNA and RNA viruses and viroid provides a new approach to virus
and virus like detection and identification. The method provides a
pure preparation of un-degraded RNA and DNA in high yield and
can be completed within 2 h. It is particularly useful for processing
large number of samples and for isolation of RNA and DNA from
small quantities of tissues. The method does not require expensive
and environmentally hazardous reagents and equipment. It can be
performed even in low technology laboratories. The amount of tissue
required by this method is ∼50–100 mg. The quantity and the quality
of the nucleic acid extracted by this method is high enough to
perform hundreds of PCR-based reactions and also to be used in
other DNA manipulation techniques such as restriction digestion,
Southern blotting and cloning.

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Journal of Virological Sciences

July

Vol. 3, Iss. 1

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