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ARS Home » Southeast Area » Fort Pierce, Florida » U.S. Horticultural Research Laboratory » Subtropical Plant Pathology Research » Research » Publications at this Location » Publication #335595

Research Project: EMERGING DISEASES OF CITRUS, VEGETABLES, AND ORNAMENTALS

Location: Subtropical Plant Pathology Research

Title: An efficient and high fidelity method for amplification, cloning and sequencing of complete tospovirus genomic RNA segments

Author
item Marshall, S - Washington State University
item Adegbola, R - Washington State University
item Adkins, Scott
item Naidu, R - Washington State University

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/29/2016
Publication Date: 1/9/2017
Citation: Marshall, S.H., Adegbola, R.O., Adkins, S.T., Naidu, R.A. 2017. An efficient and high fidelity method for amplification, cloning and sequencing of complete tospovirus genomic RNA segments. Journal of Virological Methods. 242:22-26.

Interpretive Summary: Methods were optimized for amplification of the complete M- and S-RNA segments of Tomato spotted wilt virus and Impatiens necrotic spot virus as a single fragment. This strategy ensures that all sequence determined is from a single viral RNA molecule.

Technical Abstract: Amplification and sequencing of the complete M- and S-RNA segments of Tomato spotted wilt virus and Impatiens necrotic spot virus as a single fragment is useful for whole genome sequencing of tospoviruses co-infecting a single host plant. It avoids issues associated with overlapping amplicon-based approaches and will find application for investigation of epidemiology and evolution of tospoviruses in mixed infections within natural hosts.