| | Short-range PCR amplification of coding regions followed by pooled |
| | multiplexed tagged sequencing on a GS FLX 454 instrument. Samples |
| | were amplified using primers that carried one of 16 3-base tags, to |
| | resolve sample identities of reads after pooling. Reads were mapped |
| | using BLAST, and single base substitution variants were identified by |
| | parsing the alignments. Genotypes at variant sites were determined |
| | using a Bayesian algorithm that combined read information across all |
| | sequenced individuals to estimate per-site genotype frequencies and |
| | error rates. |
| | The reported assayed sequence represents the median interval over |
| | which individuals carrying the alternate allele had sequencing reads |
| | of that allele that were otherwise in complete agreement with the |
| | genomic reference sequence, truncated to no more than 100 bp. |
