Multiplexed assembly and annotation of synthetic biology constructs using long-read Nanopore sequencing

A high-throughput plasmid sequencing approach using DNA transposition and Oxford Nanopore sequencing. The method, termed Circuit-seq, creates robust, full-length, and accurate plasmid assemblies without prior knowledge of sequences.

The approach demonstrates effectiveness across a wide range of plasmid sizes and complexities, generating full-length, contiguous plasmid maps. The technique leverages long-read data to characterize epigenetic marks and estimate contamination levels.

The work was recognized as an ACS Editors’ Choice Article Selection.

Circuit-seq scales to large numbers of samples at a lower per-sample cost than commercial Sanger sequencing, while maintaining practicality through low equipment requirements.