Figure 1.

Confirming the diversity of the molecular barcode

(A) The barcode, located in the NS4B region of the WNV genome, contains degenerate nucleotides inserted at every third codon position for 11 consecutive codons. Degenerate nucleotides produce synonymous changes and do not alter the encoded amino acid. (created with weblogo.berkeley.edu) (B) Rarefaction analysis of sequenced bcWNV stock samples estimates barcode richness as a function of sequencing depth. RNA was extracted from 50, 100, and 200ul of virus, deep sequenced and (C) barcode frequencies were determined from abundance values and ranked from most to least frequent across three stock samples and a merged population containing unique barcodes from all three stock populations (D) and number and frequency of detected barcodes was determined.