Forensic Medical Examination and Identification Center of Beijing Public Security Bureau, Beijing, PR China.
Electrophoresis. 2011 Jun;32(13):1631-8. doi: 10.1002/elps.201000694.
A common problem in the analysis of forensic DNA evidence is the presence of environmentally degraded and inhibited DNA. Such samples produce a variety of interpretational problems such as allele imbalance, allele dropout and sequence specific inhibition. In an attempt to develop methods to enhance the recovery of this type of evidence, magnetic bead hybridization has been applied to extract and preconcentrate DNA sequences containing short tandem repeat (STR) alleles of interest. In this work, genomic DNA was fragmented by heating, and sequences associated with STR alleles were selectively hybridized to allele-specific biotinylated probes. Each particular biotinylated probe-DNA complex was bound to streptavidin-coated magnetic beads using enabling enrichment of target DNA sequences. Experiments conducted using degraded DNA samples, as well as samples containing a large concentration of inhibitory substances, showed good specificity and recovery of missing alleles. Based on the favorable results obtained with these specific probes, this method should prove useful as a tool to improve the recovery of alleles from degraded and inhibited DNA samples.
在法医 DNA 证据分析中,一个常见的问题是存在环境降解和抑制的 DNA。此类样本会产生各种解释问题,例如等位基因不平衡、等位基因缺失和序列特异性抑制。为了开发增强此类证据回收的方法,已经将磁珠杂交应用于提取和预浓缩含有短串联重复 (STR) 感兴趣等位基因的 DNA 序列。在这项工作中,通过加热使基因组 DNA 碎片化,并且与 STR 等位基因相关的序列选择性地与等位基因特异性生物素化探针杂交。每个特定的生物素化探针-DNA 复合物都使用链霉亲和素包被的磁性珠结合,从而能够富集目标 DNA 序列。使用降解 DNA 样本以及含有大量抑制物质的样本进行的实验表明,该方法具有良好的特异性和缺失等位基因的回收率。基于这些特定探针获得的有利结果,该方法应该可以作为一种有用的工具,用于从降解和抑制的 DNA 样本中回收等位基因。