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High-Throughput Plasmid Sequencing Application Note | Introduction

Plasmids are ubiquitously used in recombinant DNA technology for the cloning, engineering, transfer, and expression of DNA fragments or genes. Broad-based access to cost-effective, massively parallel, next-generation sequencing (NGS), and advances in synthetic biology have revolutionized the scale on which plasmids can be constructed, screened, and manipulated for wide-ranging applications, including biomarker discovery, antibody engineering, genome editing, and gene therapy. 

NGS library preparation throughput, performance, and cost remain bottlenecks in high-throughput plasmid sequencing. In this study, we demonstrate the utility of plexWell Library Preparation Kits for high-quality, high-throughput plasmid sequencing on benchtop sequencers. 

Benefits of plexWell Library Preparation Kits for high-throughput plasmid sequencing on benchtop sequencers:

  • The unique, streamlined workflow with integrated normalization significantly reduces hands-on time and enables the processing of up to 384 plasmids by a single technician in a standard workday.
  • Effective auto-normalization results in even read distributions, without careful quantification and/or normalization of input DNA or sequencing-ready libraries.
  • Longer and tunable fragment sizes and uniform coverage enable high de novo plasmid assembly quality and success rates, from less sequencing data than required for other transposase-based workflows.
  • Plate-based reagents, flexible kit configurations, and up to 2,304 sequencing barcodes provide flexibility in terms of batch sizes, facilitate implementation in both manual and automated plasmid sequencing pipelines, and allow for optimal utilization of available sequencing capacity.

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