학술논문
Whole human exome capture for high-throughput sequencing
Document Type
Report
Source
Genome. July 1, 2010, Vol. 53 Issue 7, p568, 7 p.
Subject
Language
English
ISSN
0831-2796
Abstract
Introduction Next-generation sequencing technologies have revolutionized the ways large genomes are sequenced and have lowered the sequencing cost per nucleotide by several orders of magnitude, which is very useful in [...]
We captured the whole human exome by hybridization using synthesized oligonucleotides, based on a high-density microarray design, and we sequenced those captured human exons using high-throughput sequencing on a Genome Sequencer FLX instrument. Of the uniquely mapped reads, 71% fell within target regions, and these corresponded to coverage of 94% of human genes, 87% of exons, and 70% of the total base-pair length of the CCDS set. Our study provides strong evidence for the practical usefulness of this method on a genome-wide scale, showing the resequenced whole human exome database with 501 microRNAs and 307 novel SNPs. Key words: exon capture, next-generation sequencing, GS FLX sequencer. Les auteurs ont capture l'exome humain complet par hybridation au moyen d'oligonucleotides synthetises, base sur un design de puce d'ADN a haute densite, et ils ont sequence; ces exons captures par sequencage a haut debit sur un instrument Genome Sequencer FLX. Des sequences a position unique, 71 % provenaient des regions ciblees et celles-ci offraient une couverture de 94 % des genes humains, 87 % des exons, et 70 % de la taille du jeu CCDS mesure en paires de bases. Cette etude fournit des donnees convaincantes quant a l'utilite pratique de cette methode employee a une echelle genomique ainsi que d'une base de donnees issue du re-sequencage du genome humain comprenant 501 microARN et 307 nouveaux SNP. Mots-cles: capture d'exons, sequencage de seconde generation, sequenceur GS FLX. [Traduit par la Redaction]
We captured the whole human exome by hybridization using synthesized oligonucleotides, based on a high-density microarray design, and we sequenced those captured human exons using high-throughput sequencing on a Genome Sequencer FLX instrument. Of the uniquely mapped reads, 71% fell within target regions, and these corresponded to coverage of 94% of human genes, 87% of exons, and 70% of the total base-pair length of the CCDS set. Our study provides strong evidence for the practical usefulness of this method on a genome-wide scale, showing the resequenced whole human exome database with 501 microRNAs and 307 novel SNPs. Key words: exon capture, next-generation sequencing, GS FLX sequencer. Les auteurs ont capture l'exome humain complet par hybridation au moyen d'oligonucleotides synthetises, base sur un design de puce d'ADN a haute densite, et ils ont sequence; ces exons captures par sequencage a haut debit sur un instrument Genome Sequencer FLX. Des sequences a position unique, 71 % provenaient des regions ciblees et celles-ci offraient une couverture de 94 % des genes humains, 87 % des exons, et 70 % de la taille du jeu CCDS mesure en paires de bases. Cette etude fournit des donnees convaincantes quant a l'utilite pratique de cette methode employee a une echelle genomique ainsi que d'une base de donnees issue du re-sequencage du genome humain comprenant 501 microARN et 307 nouveaux SNP. Mots-cles: capture d'exons, sequencage de seconde generation, sequenceur GS FLX. [Traduit par la Redaction]