- Chen, Zhoutao;
- Pham, Long;
- Wu, Tsai-Chin;
- Mo, Guoya;
- Xia, Yu;
- Chang, Peter L;
- Porter, Devin;
- Phan, Tan;
- Che, Huu;
- Tran, Hao;
- Bansal, Vikas;
- Shaffer, Justin;
- Belda-Ferre, Pedro;
- Humphrey, Greg;
- Knight, Rob;
- Pevzner, Pavel;
- Pham, Son;
- Wang, Yong;
- Lei, Ming
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 requirements. We have developed a single-tube Transposase Enzyme Linked Long-read Sequencing (TELL-seq) technology, which enables a low-cost, high-accuracy, and high-throughput short-read second-generation sequencer to generate over 100 kb of long-range sequencing information with as little 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 barcoded linked-reads are used to successfully assemble genomes ranging from microbes to human. These linked-reads also generate megabase-long phased blocks and provide a cost-effective tool for detecting structural variants in a genome, which are important to identify compound heterozygosity in recessive Mendelian diseases and discover genetic drivers and diagnostic biomarkers in cancers.