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A strategy was developed for sequence-independent synthesis and amplification of full-length cDNA of 3-4 kb genes of dsRNA viruses. The method of single primer amplification (Lambden et al., 1992) was adapted by the inclusion of a 3' poly(A) tail to an oligonucleotide ligated to dsRNA genome segments as a template for oligo(dT)-primed cDNA synthesis. Full-length copies of the largest genome segments, 1 (4 kb) and 2 (3 kb), of African horse sickness virus (AHSV) have been cloned, terminally sequenced and expressed in vitro.

Type

Journal article

Journal

J Virol Methods

Publication Date

06/1998

Volume

72

Pages

243 - 247

Keywords

African Horse Sickness Virus, Animals, Blotting, Northern, DNA Primers, DNA, Complementary, Electrophoresis, Agar Gel, Gene Amplification, Genome, Viral, Horses, RNA Viruses