New developments in high‐throughput resequencing and variation detection using high density microarrays
New developments in high‐throughput resequencing and variation detection using high density microarrays
复制标题
使用高密度微阵列进行高通量重测序和变异检测的新进展
作者：
J. Warrington;Nila A. Shah;Xiyin Chen;M. Janis;Chunmei Liu;S. Kondapalli;Vivian Reyes;M. P. Savage;Zhaomei Zhang;Richard Watts;Maria deGuzman;A. Berno;Jim Snyder;J. Baid
我们开发了一种利用高密度微阵列进行单核苷酸多态(SNP)发现的高通量重测序方法。在这项研究的两年过程中，在样品准备方法、杂交分析、阵列处理和分析方法方面进行了一些改进。来自三个不同种族的40个无关个体的DNA被扩增、标记，并与用代表基因组、编码和调节区的探针设计的阵列杂交。包括长聚合酶链式反应和半自动的使用在内的技术改进降低了33%的标记和片段化成本。自动化方面的改进包括开发用于阵列的扫描仪自动装载机、更快的阵列洗涤站以及链接的实验室跟踪和数据管理系统。验证较小的特征尺寸，20×24微米，允许在每个微阵列上同时筛选30kb的正义DNA和30kb的反义DNA，将吞吐量提高到每两名实验室人员每天1.4Mb。使用高密度重测序和变异检测阵列(微阵列)，在8.3Mb的人类基因组中发现了超过15,000个SNPs。Hum Mutat 19：402-409,2002。©2002 Wiley-Liss公司
We developed a high‐throughput method for resequencing for single nucleotide polymorphism (SNP) discovery using high‐density microarrays. Over the two‐year course of this study a number of improvements in sample preparation methods, hybridization assay, array handling, and analysis method were developed and implemented. DNA from 40 unrelated individuals of three different ethnic origins was amplified, labeled, and hybridized to arrays designed with probes representing genomic, coding, and regulatory regions. Protocol improvements including the use of long PCR and semi‐automation reduced labeling and fragmentation costs by 33%. Automation improvements include the development of a scanner autoloader for arrays, a faster array wash station, and a linked laboratory tracking and data management system. Validation of a smaller feature size, 20 × 24 microns, allowed the simultaneous screening of 30‐kb sense and 30‐kb antisense DNA on each microarray, increasing throughput to 1.4 Mb per day per two laboratory personnel. More than 15,000 SNPs were identified in 8.3 Mb of the human genome using high‐density resequencing and variation detection arrays (microarrays). Hum Mutat 19:402–409, 2002. © 2002 Wiley‐Liss, Inc.
影响因子：
56.9
作者：
Wang, DG;Fan, JB;Lander, ES
通讯作者：
Lander, ES