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Author(s): Ekholm, Jenny and Gu, Jenny and Burke, Jeff and Kim, Michelle, and Sibert, Justin and Liu, Klvin

We have streamlined the SMRTbell library generation protocols with improved workflows to deliver seamless end-to-end solutions from sample to analysis. A key improvement is the development of a single-tube reaction strategy that shortened hands-on time needed to generate each SMRTbell library, reduced time-consuming AM Pure purification steps, and minimized sample-handling induced gDNA damage to improve the integrity of long-insert SMRTbell templates for sequencing. The improved protocols support all large-insert genomic libraries, multiplexed microbial genomes, and amplicon sequencing. These advances enable completion of library preparation in less than a day (approximately 4 hours) and opens opportunities for automated library preparation for large-scale projects. Here we share data summarizing performance of the new SMRTbell Express Template Kit 2.0 representing our solutions for 10 kb and >50 kb large-insert genomic libraries, complete microbial genome assemblies, and high-throughput amplicon sequencing. The improved throughput of the Sequel System with read lengths up to 30 kb and high consensus accuracy (> 99.999% accuracy) makes sequencing with high-quality results increasingly assessible to the community.

Organization: PacBio
Year: 2019

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