Ultralow-input single-tube linked-read library method enables short-read second-generation sequencing systems to routinely generate highly accurate and economical long-range sequencing information
| Autor: | Pedro Belda-Ferre, Long Pham, Yu Xia, Guoya Mo, Zhoutao Chen, Huu Che, Yong Wang, Justin P. Shaffer, Tan Phan, Vikas Bansal, Peter L. Chang, Devin Porter, Wu Tsai-Chin, Ming Lei, Rob Knight, Hao Tran, Greg Humphrey, Son Pham, Pavel A. Pevzner |
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| Rok vydání: | 2020 |
| Předmět: |
Sequence analysis
Bioinformatics Sequence assembly Computational biology Biology Barcode Genome Medical and Health Sciences law.invention Workflow Structural variation 03 medical and health sciences 0302 clinical medicine law HLA Antigens Genetics DNA Barcoding Taxonomic Humans Genomic library Genetics (clinical) Transposase 030304 developmental biology Gene Library 0303 health sciences Genome Human Human Genome Computational Biology Genetic Variation Taxonomic High-Throughput Nucleotide Sequencing Sequence Analysis DNA DNA Genomics Biological Sciences Short read DNA Barcoding Haplotypes Generic health relevance Erratum Sequence Analysis 030217 neurology & neurosurgery Human |
| Zdroj: | Genome research, vol 30, iss 6 Genome Res |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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