Microarray-based STR genotyping using RecA-mediated ligation.
Microarray-based STR genotyping using RecA-mediated ligation.
复制标题
使用 RecA 介导的连接进行基于微阵列的 STR 基因分型。
DOI：
10.1093/nar/gkq657
复制
发表时间：
2010
影响因子：
14.9
通讯作者：
Wagner,Robert
中科院分区：
文献类型：
--
作者：
Herrmann,David;Rose,Emily;Muller,Uwe;Wagner,Robert
我们描述了一种新的检测能够准确地确定短串联重复序列（STR）等位基因的长度。STR基因分型是利用RecA介导的连接（RML），它结合了高保真的RecA介导的同源性搜索与等位基因特异性连接。RecA催化合成寡核苷酸与双链DNA靶的一条链（在这种情况下为PCR扩增子）的配对。只有当两个相邻的寡核苷酸与STR区域碱基配对而没有任何重叠或缺口时，才发生连接。需要RecA活性来克服在寄存器中退火重复序列的固有困难。该测定能够确定人类样品的STR基因型，易于适应于高通量或自动化系统，并且可以在诊断和法医应用中具有广泛的实用性。
We describe a novel assay capable of accurately determining the length of short tandem repeat (STR) alleles. STR genotyping is achieved utilizing RecA-mediated ligation (RML), which combines the high fidelity of RecA-mediated homology searching with allele-specific ligation. RecA catalyzes the pairing of synthetic oligonucleotides with one strand of a double-stranded DNA target, in this case a PCR amplicon. Ligation occurs only when two adjacent oligonucleotides are base paired to the STR region without any overlap or gap. RecA activity is required to overcome the inherent difficulty of annealing repeated sequences in register. This assay is capable of determining STR genotypes of human samples, is easily adapted to high throughput or automated systems and can have widespread utility in diagnostic and forensic applications.