断裂位点批量映射，一种用于分析和鉴定哺乳动物细胞中DNA断裂位点的快速方法。
Breaksite batch mapping, a rapid method for assay and identification of DNA breaksites in mammalian cells.
作者信息
Kong Q, Maizels N
机构信息
Department of Molecular Biophysics and Biochemistry and Department of Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520-8024, USA.
出版信息
Nucleic Acids Res. 2001 Mar 15;29(6):E33. doi: 10.1093/nar/29.6.e33.
DNA breaks occur during many processes in mammalian cells, including recombination, repair, mutagenesis and apoptosis. Here we report a simple and rapid method for assaying DNA breaks and identifying DNA breaksites. Breaksites are first tagged and amplified by ligation-mediated PCR (LM-PCR), using nested PCR primers to increase the specificity and sensitivity of amplification. Breaksites are then mapped by batch sequencing LM-PCR products. This allows easy identification of multiple breaksites per reaction without tedious fractionation of PCR products by gel electrophoresis or cloning. Breaksite batch mapping requires little starting material and can be used to identify either single- or double-strand breaks.
DNA断裂发生在哺乳动物细胞的许多过程中，包括重组、修复、诱变和凋亡。在此，我们报告一种简单快速的方法来检测DNA断裂并识别DNA断裂位点。首先通过连接介导的PCR（LM-PCR）对断裂位点进行标记和扩增，使用巢式PCR引物来提高扩增的特异性和灵敏度。然后通过对LM-PCR产物进行批量测序来定位断裂位点。这使得每个反应能够轻松识别多个断裂位点，而无需通过凝胶电泳或克隆对PCR产物进行繁琐的分级分离。断裂位点批量定位所需起始材料很少，可用于识别单链或双链断裂。
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