Determination of coronaviral poly(A) tail length in virions used for inoculation.

<p>(A) Strategy for determining coronaviral poly(A) tail length in virus used for inoculation. RNA extracted from BCoV harvested at 48 hpi was decapped and head-to-tail ligated. BCoV 5’ UTR-positive-strand-specific primer 2 (for RT) and BCoV 3’ UTR-negative-strand specific primer 1 were used for RT-PCR with the ligated RNA as template. Note that the head-to-tail ligation could be inter-molecular or intra-molecular. Inter-molecular-ligated RNA is represented here and in the following figures. The amplified RT-PCR product was sequenced to determine poly(A) tail length. (B) Sequence of the amplified RT-PCR product. The poly(A) tail length in virus used for inoculation was ~45 nt.