Homo sapiens isolate:HD701 Raw sequence reads. Homo sapiens isolate:HD701

Single molecule sequencing (SMS) platforms enable base sequences to be read directly from individual strands of DNA in real-time. Though capable of long read lengths, SMS platforms currently suffer from low throughput compared to competing short-read sequencing platforms. Here, we present a novel strategy for sequencing library preparation, dubbed ConcatSeq, which increases the throughput of SMS platforms by generating long concatenated templates from pools of short DNA molecules. We show adaptation of this technique to two target enrichment workflows, commonly used for oncology applications, and demonstrate feasibility using PacBio single molecule real-time (SMRT) Technology.