Abi prism 3140

Amplicons were analyzed using agarose gel electrophoresis. Depending on the
presence of a single or multiple bands on the gel, amplicons were either
directly purified with QIAquick PCR Purification kit or gel-isolated and
purified using QIAquick Gel Extraction kit (Qiagen, Courtaboeuf, France)
following the manufacturer’s protocol.
Resulting amplicons were subjected to direct sequencing using the BigDye
terminator v3.1 kit (Applied Biosystems) and an ABI Prism 3140 automated
sequencer (Applied Biosystems). The sequencing of each amplicon was performed in
both directions using semi-nested PCR primers. Sequence data obtained using the
ABI PRISM kits were viewed, assembled and edited using the CLC Main Workbench
5.7.2 software (CLC bio, Aarhus, Denmark). Consensus sequences were submitted to
databases with the following accession numbers: MK310554 to MK310568 and
MK310788 to MK310984 respectively for the VP1 and VP3-VP1 sequences of NPEVs;
MK310569 to K310641, MK310642 to MK310714 and MK310715 to MK310787 respectively
for the 2C^ATPase, 3D^pol and 5’UTR sequences of cVDPVs;
MK310985 to MK311038 for the full-length VP1 sequences of cVDPVs; MK311039 to
MK311132 for the full-length VP1 sequences of Sabin-like PVs.

Amplicons were purified using the QIAquick PCR Purification kit (Qiagen, Courtaboeuf, France) following the manufacturer’s protocol. Purified amplicons were subjected to direct double strands sequencing using nested PCR primers, the BigDye terminator v3.1 kit (Applied Biosystems) and the ABI Prism 3140 automated sequencer (Applied Biosystems).
Consensus sequence editing, multiple sequences alignments and pairwise sequence comparisons were carried out with the CLC Main Workbench 5.7.2 software (CLC bio, Aarhus, Denmark).
GenBank accession numbers for the nucleocapsid gene sequences of RABV determined in this study have been assigned as MF537505 to MF537580.