Fig 5. Sequencing of ZIKV-BC-1.0 and ZIKV-IC isolated from nonpregnant rhesus macaques.

ZIKV RNA was isolated from plasma at the indicated time points from each of the animals infected with A.) ZIKV-IC or B.) ZIKV-BC-1.0. Viral RNA was reverse transcribed and then multiplex PCR was performed as described in materials and methods. PCR products were tagged and sequenced. A.) The sequence mapping to the region containing the molecular barcode was interrogated for ZIKV-IC, and the frequency of wild type and non-wild type ZIKV sequences are shown. The theoretical number of cDNA molecules used in each PCR reaction is shown in Table 2. Each sample was sequenced in duplicate, as labeled by A and B. B.) The frequency of each barcode in the population is shown for ZIKV-BC-1.0 and the nonpregnant animals infected with ZIKV-BC-1.0. The frequency of the wild type sequence in the region of the barcode is shown as Zika_WT. The frequency of any sequence in the region of the barcode that was not considered authentic is listed as ‘Other.’ C.) The number of authentic barcodes detected in the three nonpregnant animals and the stock were counted. The data for each individual replicate are shown.