Nonradioactive, photobiotin-labelled DNA probes for the routine diagnosis of barley yellow dwarf virus.

Abstract:

:Photobiotin was used to prepare biotinylated, nonradioactive nucleic acid probes for the detection of the RNA of barley yellow dwarf virus (BYDV) in plant extracts. A 1.7-kb cDNA of the PAV isolate of BYDV in the plasmid pUC8 vector was biotinylated and then used intact or as sonicated double-stranded DNA fragments. Simple methods were developed for the preparation of partially purified nucleic acid extracts of cereals and their spotting, after formaldehyde treatment, onto nitrocellulose membranes. After hybridization, biotin-labelled DNA bound to BYDV RNA on the nitrocellulose was detected with an avidin-alkaline phosphatase conjugate. BYDV RNA was readily detected with a sensitivity similar to that found with the same probe labelled with 32P by nick translation. Healthy plant extracts gave colourless spots.

journal_name

J Virol Methods

authors

Habili N,McInnes JL,Symons RH

doi

10.1016/0166-0934(87)90007-3

subject

Has Abstract

pub_date

1987-06-01 00:00:00

pages

225-37

issue

3

eissn

0166-0934

issn

1879-0984

pii

0166-0934(87)90007-3

journal_volume

16

pub_type

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