Fig. 1.

Outline of RT-PCR strategies that have been used to amplify the complete genome sequences of FMDV. (a) Long overlapping products (∼3 kb) are generated by PCR using full-length cDNA as a template. Sequences are obtained using a panel of specific sequencing primers (see ref. 3). (b) Short products (∼700 bp) are generated using FMDV-specific primers, which also incorporate regions (labelled F and R) targeted by the sequencing primers (10). (c) Long-range RT-PCR is used to amplify a product comprising the complete L fragment of FMDV. This may either be sequenced using many specific primers (11) or can be fragmented by restriction digest and cloned into a bacterial plasmid vector (pilot studies using this method have been undertaken by IAH in collaboration with the Wellcome Trust Sanger Institute, Cambridge).