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Sequence-independent amplification and cloning of large dsRNA virus genome segments by poly(dA)-oligonucleotide
F T Vreede1, M Cloete, G B Napier
1Onderstepoort Veterinary Institute, South Africa. frank@moon.ovi.ac.za
Journal of Virological Methods
|August 7, 1998
Abstract:
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.