High throughput DNA sequence variant detection by conformation sensitive capillary electrophoresis and automated peak comparison
High throughput DNA sequence variant detection by conformation sensitive capillary electrophoresis and automated peak comparison
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DOI：
10.1016/j.ygeno.2005.11.008
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发表时间：
2006-03-01
期刊：
影响因子：
4.4
通讯作者：
Wooster, R
中科院分区：
文献类型：
--
作者：
Davies, H;Dicks, E;Wooster, R
我们报道了一种利用多毛细管自动测序仪的基于异源双链的突变检测方法的开发，这种方法被称为构象敏感毛细管电泳（CSCE）。我们优化后的CSCE方案检测到了95个已知碱基替换序列变体中的93个。自从该方法优化以来，我们已经分析了2.15亿碱基对的DNA，并鉴定出3397个独特的变体。对该数据集的分析表明，在平均PCR产物的中间56%部分，CSCE的灵敏度高于95%。为了充分利用这种方法的突变检测能力，我们开发了软件canplot，它能自动比较正常结果和测试结果，以便对最有可能包含变体的样本进行优先级排序。利用多种荧光染料，CSCE每天能够在一台ABI3730上筛选超过220万碱基对。因此，这种技术适用于需要快速且灵敏的DNA突变检测系统的项目。（c）2005爱思唯尔公司。保留所有权利。
We report the development of a heteroduplex-based mutation detection method using multicapillary automated sequencers, known as conformation-sensitive capillary electrophoresis (CSCE). Our optimized CSCE protocol detected 93 of 95 known base substitution sequence variants. Since the optimization of the method, we have analyzed 215 Mb of DNA and identified 3397 unique variants. An analysis of this data set indicates that the sensitivity of CSCE is above 95% in the central 56% of the average PCR product. To fully exploit the mutation detection capacity of this method, we have developed software, canplot, which automatically compares normal and test results to prioritize samples that are most likely to contain variants. Using multiple fluorescent dyes, CSCE has the capacity to screen over 2.2 Mb on one ABI3730 each day. Therefore this technique is suitable for projects where a rapid and sensitive DNA mutation detection system is required. (c) 2005 Elsevier Inc. All rights reserved.