Fig. 4
Proposed stratified approach for mutation screening by HaloplexHS target enrichment and Illumina HiSeq4000 sequencing according to DNA quality. The DNA samples that support PCR of ≥400 bp genomic fragment are amenable for mutation analysis with a single replicate, with only variants at 15–20 AAD required for further validation, while those with suboptimal quality (PCR of up to 300 bp) are better analysed in duplicate with reproducible variants >15 AAD regarded as true genetic changes. DNA samples that support PCR of only 200 bp genomic fragment are not suitable for targeted sequencing under the conditions specified.
