Detection of CCND1 gene copy number variations using multiplex ligation-dependent probe amplification and fluorescence in situ hybridization Methods.
Detection of CCND1 gene copy number variations using multiplex ligation-dependent probe amplification and fluorescence in situ hybridization Methods.
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使用多重连接依赖性探针扩增和荧光原位杂交方法检测 CCND1 基因拷贝数变异。
DOI：
10.1007/978-1-4939-7565-5_10
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Ooi A and Oyama T
中科院分区：
文献类型：
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作者：
Ooi A;Inokuchi M;Horike S;Kawashima H;Ishikawa S;Ikeda H;Nakamura R;Oyama T;Dobashi Y;Ooi A and Oyama T
CCND1 位点位于 11q13，编码 G1-S 调节蛋白，细胞周期蛋白 D1。 Cyclin D1 在各种类型的癌症中经常被扩增，是一个有吸引力的潜在治疗靶点。多重连接依赖性探针扩增 (MLPA) 是一种新的高分辨率方法，用于在单个反应中检测来自福尔马林固定、石蜡包埋材料的少量 DNA 片段中包括 CCND1 在内的多种基因的扩增。然而，这种方法基于 PCR 并对许多不同的细胞进行平均，因此理论上必须通过荧光原位杂交 (FISH) 等形态学方法进行验证。在这里，我们描述了通过市售的 MLPA 试剂盒和使用细菌人工染色体 (BAC) 探针的 FISH 检测 CCND1 基因拷贝数变异。
TheCCND1locus is located in 11q13 and encodes the G1–S regulatory protein, cyclin D1. Cyclin D1 is frequently amplified in various types of cancers, and is an attractive potential therapeutic target. Multiplex ligation-dependent probe amplification (MLPA) is a new, high-resolution method for the detection of amplification of numerous genes includingCCND1in small amounts of DNA fragments derived from formalin-fixed, paraffin-embedded material in a single reaction. This approach is, however, based on PCR and averages many different cells, so validation by morphological methods such as fluorescence in situ hybridization (FISH) is theoretically mandatory. Here we describe detection ofCCND1gene copy number variations by commercially available MLPA kits and FISH using a bacterial artificial chromosome (BAC) probe.