Figure 1

Probe construction, target capture and reciprocal paired-end sequencing. (a) Each cLPP contains a common linker flanked by post-capture amplification sites (red and green) and two target-specific capturing arms (blue and orange). Probe ends are trimmed (BsaI and MlyI) and 5'-phosphorylated to produce functional cLPPs. (b) Multiplex probe-target hybridization followed by gap-filling and ligation triggers probe circularization and target capture. (c) Capture libraries are multiplex-amplified using hybrid primers that anneal to the probes' amplification sites and add Illumina sequencing adaptors (P5 or P7) and sample-specific barcodes. This is done in two separate PCRs during which the adaptors swap positions at the ends of templates. Both PCRs are pooled for reciprocal PE sequencing of both DNA strands.