Ultra-low input single tube linked read library method enables short read NGS systems to generate highly accurate and economical long-range sequencing information for de novo assembly and haplotype phasing

Long range sequencing information is required for haplotype phasing, de novo assembly and structural variation detection. Current long read sequencing technologies can provide valuable long-range information but at a high cost with low accuracy and high DNA input requirement. We developed a single tube Transposase Enzyme Linked Long-read Sequencing (TELL-Seq) technology which enables a relatively low cost, high accuracy and high throughput short read next generation sequencer to routinely generate over 100 Kb long range sequencing information using as low as 0.1 ng input material. In a PCR tube, millions of clonally barcoded beads are used to uniquely barcode long DNA molecules in an open bulk reaction without dilution and compartmentation. The barcode linked reads are used to successfully assemble genomes ranging from microbes to human. These linked reads also generate mega-base long phased blocks and provide a cost-effective tool for detecting structural variants in the genome.