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ARS Home » Pacific West Area » Corvallis, Oregon » Horticultural Crops Research Unit » Research » Publications at this Location » Publication #209636

Title: A New Protocol for Extraction of Double-Stranded RNA from Plants

Author
item Martin, Robert
item TZANETAKIS, I - OREGON STATE UNIVERSITY

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: 4/25/2007
Publication Date: 6/1/2007
Citation: Martin, R.R., Tzanetakis, I.E. 2007. A new protocol for extraction of double-stranded RNA from plants. Phytopathology. 97. http://meeting.apsnet.org/cfa/ssPresentationView.cfm?PresentationID=2485&

Interpretive Summary:

Technical Abstract: There are several unidentified virus-like diseases of blueberry that have been recalcitrant to standard methods of virus characterization based on double-stranded RNA (dsRNA). Relatives of these viruses yield large amounts of dsRNA in hosts other than blueberry. Modifications in dsRNA extraction protocols, including high molarity salts, increased buffering capacity and pH of the extraction buffers failed to improve the yield of dsRNA from blueberry. A new protocol based on a lithium salt and detergent extraction buffer that does not use any organic solvent other than ethanol has been developed. The protocol uses half the amount of tissue suggested in the other protocols, works effectively with blueberry tissue and yields sufficient amounts of dsRNA for detection in EtBr stained agarose gels and for cloning. The protocol has been used on several small fruit hosts and the yield of dsRNA was similar to previously described protocols.