An efficient, tunable solution for pre-capture multiplexing in targeted sequencing via novel chemistry and semiconductor DNA synthesis technology
posterposted on 20.04.2020, 22:42 by Jiashi Wang, Cheng Song, Raymond Miller, Jianpeng Wang, Lumeng Ye, Hong Li, Heidi Huang
Pre-capture multiplexing is a cost-effective and efficient way for targeted next-generation sequencing. To allow high-throughput pre-capture pooling without compromising the sequencing quality, we developed a series of reagents, including 384+ plex customizable adaptors, TRUE universal blockers, and a fast and cost-effective hybrid capture solution using semiconductor DNA synthesis technology. Our data shows our adaptors have an extremely lost cross-contamination rate, which can minimize the index hopping during pre-capture pooling. The TRUE universal blockers using unique chemistry can bind to adapters regardless of the index length, simplify the workflow and still yield consistent capture efficiency during sample multiplexing. Also, the GenFisher capture probes, produced by semiconductor DNA synthesis technology, can deliver high capture efficiency after pre-capture pooling, with the potential of evolution via the test-learn cycle.