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Scheme of primer design for multiplex MB-NGS.
Eleven sets of primers were designed to cover ESR1-LBD (the light gray box, c.928-1641), and their target regions were indicated by arrows. The two dark gray boxes showed the gaps not covered by these primers, and the coverage was 96.8% (691 / 714 bp). The hatching box represents the hotspot region (c.1606-1614) of ESR1 mutations.
