Design and Generation of MLPA Probe Sets for Combined Copy Number and Small-Mutation Analysis of Human Genes: EGFR as an Example
Design and Generation of MLPA Probe Sets for Combined Copy Number and Small-Mutation Analysis of Human Genes: EGFR as an Example
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DOI：
10.1100/tsw.2010.195
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发表时间：
2010-01-01
影响因子：
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通讯作者：
Kozlowski, Piotr
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文献类型：
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作者：
Marcinkowska, Malgorzata;Wong, Kwok-Kin;Kozlowski, Piotr
多重连接依赖性探针扩增（MLPA）是一种多重拷贝数分析方法，通常用于许多临床和研究实验室中识别大突变。标准MLPA设置的最重要缺点之一是生成长MLPA探针的过程复杂且因此昂贵。该缺点基本上限制了MLPA对那些可用的商业试剂盒的基因组区域的适用性。在这里，我们提出了一个简单的协议，设计MLPA探针集，完全由短的寡核苷酸半探针通过化学合成。作为一个例子，我们提出了设计和生成的MLPA检测平行拷贝数和EGFR基因的小突变分析。
Multiplex ligation-dependent probe amplification (MLPA) is a multiplex copy number analysis method that is routinely used to identify large mutations in many clinical and research labs. One of the most important drawbacks of the standard MLPA setup is a complicated, and therefore expensive, procedure of generating long MLPA probes. This drawback substantially limits the applicability of MLPA to those genomic regions for which ready-to-use commercial kits are available. Here we present a simple protocol for designing MLPA probe sets that are composed entirely of short oligonucleotide half-probes generated through chemical synthesis. As an example, we present the design and generation of an MLPA assay for parallel copy number and small-mutation analysis of the EGFR gene.