Hebei Key Laboratory of Forensic Medicine, Department of Forensic Medicine, Hebei Medical University, Shijiazhuang, Hebei, People's Republic of China.
Mol Biol Rep. 2012 Jun;39(6):6949-57. doi: 10.1007/s11033-012-1522-2. Epub 2012 Feb 5.
We report a pyrosequencing method for detecting a short amelogenin fragment to aid the gender identification. The PCR products (44/45 bp), including primers and target sequence (4/5 bp) consisting of three point mutations and one indel mutation, were sequenced by the pyrosequencing method. 100 randomly chosen DNA samples of healthy donors were analyzed with this method, and all of them were correctly typed. The sensitivity of the technique was 0.5 ng template DNA. No specific peak was found in any detected animals or organisms except for monkey. For blood samples that were left outside for 26 weeks and DNA degraded artificially by digesting with DNaseI, this method gave more accurate results than the conventional method. Moreover, four bone samples analyzed using the method gave clear pyrograph. This method is easy, quick, cheap and suitable for high-throughput analysis, especially for identifying the gender of highly-degraded DNA samples.
我们报告了一种焦磷酸测序方法，用于检测短的牙釉蛋白片段，以辅助性别鉴定。PCR 产物（44/45 bp），包括引物和目标序列（4/5 bp），由三个点突变和一个插入缺失突变组成，通过焦磷酸测序法进行测序。用该方法分析了 100 个随机选择的健康供体 DNA 样本，所有样本均被正确分型。该技术的灵敏度为 0.5 ng 模板 DNA。除了猴子，在任何检测到的动物或生物体中都没有发现特定的峰。对于在体外放置 26 周且 DNA 被 DNaseI 消化人工降解的血液样本，该方法比传统方法给出了更准确的结果。此外，用该方法分析的四个骨样本给出了清晰的图谱。该方法简单、快速、便宜，适合高通量分析，特别是用于鉴定高度降解的 DNA 样本的性别。
