A modified procedure for isolating double-stranded RNA: Application to diagnosis of Amasya cherry disease

Abstract

A novel method of isolating double-stranded RNA (dsRNA) from cherry leaves affected with Amasya cherry disease (ACD) using silica capture in combination with non-ionic cellulose was developed. Unfractionated RNAs prepared using the new procedure and used to isolate dsRNA was cleaner in terms of contamination with proteins and phenolics when compared with preparations isolated with a phenol/chloroform method. The generated dsRNA was successfully utilized in a reverse transcription-polymerase chain reaction method developed to detect a diagnostic, mycovirus dsRNA element always associated with ACD.