(42) The Sanger method
is preferred for several reasons.
Although Rothberg and his colleagues are the first to bring to market a contender to the Sanger method
, many other groups are developing challengers.
Captured sequence libraries were analyzed with both the GS-FLX and the GAII, and a subset of targeted exons was sequenced by the Sanger method
to validate the NGS-identified variants.
This sequence was verified by using back-to-back specific primers, and the amplicon was cloned and sequenced according to the Sanger method
We confirmed the AGV2 specificity of these PCR-amplified bands by direct sequencing using the Sanger method
. The sequence assay showed that the 12 sequences identified here had 98.3%-100% homology to each other and 92.2%-99.1% aa identity to AGV2 samples previously deposited in GenBank (see Figure legend for accession numbers).
In this version of the approach, following bisulphite modification, we'd use PCR with primers flanking the methylation site of interest to generate sufficient amounts of template for sequencing by either Sanger methods