Introducing a new and improved approach for verifying complete plasmid constructs

Oxford Nanopore is pleased to introduce the plasmid sequencing workflow, a new and improved approach for verifying complete plasmid constructs in your lab.

Plasmid constructs serve as fundamental tools in molecular biology labs and are used for a wide range of applications, such as protein production, genome engineering, and gene therapy. Plasmid sequence verification is a critical quality control step prior to downstream applications. However, typical short-read DNA sequencing methods only verify a partial region of the plasmid and therefore miss important information that can result in failed experiments, unnecessary troubleshooting, and increased costs.

Introducing an end-to-end solution

The new plasmid sequencing workflow is a simple, end-to-end solution that:

• generates highly accurate sequence information for the entire plasmid
• provides a simple point-and-click data analysis and report
• enables rapid, same-day turnaround time at a competitive price
• facilitates sequencing in your own lab, ensuring data remains secure.

Using this method, the entire plasmid can be sequenced rapidly, in-house at a competitive price. This enables several advantages over short-read sequencing methods. Users can:

• easily identify if insert is the correct size and orientation
• verify if the plasmid background is free of mutations
• resolve repeat regions, dimers and deletions
• spot contaminants or unexpected products
• avoid complicated primer walking and annealing issues.

After culturing colonies and performing plasmid preparation using standard 3rd party methods, samples can be prepared for sequencing using the Oxford Nanopore Rapid Barcoding Kit. The samples are then sequenced on MinION or GridION devices, and the basecalled data can be analysed using either the Clone Validation workflow in EPI2ME, or a run locally on a computer in EPI2ME Labs.

Access the workflow today

The workflow is now available in the resource centre, including an easy-to-use clone validation analysis method in EPI2ME. Users can also find more information about the workflow in this video.