High-throughput sequencing, also known as "Next-generation" sequencing technology, enables sequencing and general reading of hundreds of thousands to millions of DNA molecules in parallel at a time. The shorter length is the sign.
Technical application
Sequencing technology advances the development of scientific research. With the rapid development of second-generation sequencing technology, the scientific community has begun to use second-generation sequencing technology to solve biological problems. For example, de novo sequence of a species that has no reference sequence at the genomic level, obtaining a reference sequence for the species, laying the foundation for subsequent research and molecular breeding; whole genome resequencing of species with reference sequences ( Resequencing), scanning and detecting mutation sites at the genome-wide level, and discovering the molecular basis of individual differences. Whole transcriptome resequencing at the transcriptome level for alternative splicing, coding sequence single nucleotide polymorphism (cSNP) studies, or small RNA sequencing RNA molecules of a specific size are isolated for sequencing to discover new microRNA molecules. At the transcriptome level, combined with chromatin immunoprecipitation (ChIP) and methylated DNA immunoprecipitation (MeDIP) to detect DNA regions and genomic methylation sites that bind to specific transcription factors.
What needs to be specifically pointed out here is the application of second-generation sequencing combined with microarray technology - Targeted Resequencing. The technology first uses microarray technology to synthesize a large number of oligonucleotide probes that are capable of complementary binding to specific regions of the genome, enriching into specific segments, and then using second-generation sequencing techniques. These segments were sequenced. Agilent and Nimblegen are currently available for sequence capture, and the most widely used is human exome capture sequencing. Scientists now believe that exome sequencing is more advantageous than whole-genome resequencing, not only because of lower costs, but also because the data analysis of exome sequencing is less computationally intensive and more directly integrated with biological phenotypes.
Currently, high-throughput sequencing is beginning to be widely used to find candidate genes for disease. Researchers at the University of Nijmegen used this method to identify pathogenic mutations in Schinzel-Giedion syndrome, a rare disease that causes severe mental retardation, high tumors, and multiple congenital malformations. They sequenced the exome of four patients using Agilent SureSelect sequence capture and SOLiD with an average coverage of 43-fold and a read length of 50 nt, with each individual producing 2.7-3 GB of mapable sequence data. They focused on the 12 genes of all four patients carrying variants, eventually narrowing the candidate gene to one. Baylor College of Medicine's Genome Sequencing Center also plans to conduct research on 15 diseases in Science's top ten scientific breakthroughs, including brain cancer, liver cancer, pancreatic cancer, colon cancer, ovarian cancer, bladder cancer, heart disease, diabetes, and autism. And other genetic diseases to better understand the pathogenic mutations and the effects of mutations on the disease. Exomes have just been included in the selection that has just ended.
Significance
The birth of sequencing technology can be said to be a landmark event in the field of genomics research. This technology has led to a sharp decline in the cost of single-base sequencing of nucleic acid sequencing compared to first-generation sequencing technologies. For example, in the human genome sequencing, the human genome project at the end of the last century cost $3 billion to decode human life codes, while second-generation sequencing The human genome has been sequenced into the 10,000-year genome era. Such low cost of single base sequencing allows us to implement genome projects for more species to decipher the genomic genetic code of more biological species. At the same time, large-scale whole-genome resequencing of other species of the species has become possible in species that have completed genome sequencing.
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