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PROCESS FOR ANIMAL SPECIES IDENTIFICATION IN SAMPLES WITH GENETIC MATERIAL BASED ON MITOCHONDRIAL DNA SIZE VARIATION

机译:基于线粒体DNA大小变异的遗传材料样品动物物种鉴定过程

摘要

The present invention refers to a process for animal species identification in biological samples with genetic material of animal origin, based on variations in the size of mitochondrial 12s and 16s ribosomal RNA gene sequences. The method of this invention comprises a panel of oligonucleotide PCR (Polymerase Chain Reaction) primers, labelled with fluorescent dyes, for multiplex PCR amplification of seven mitochondrial DNA regions with insertiondeletion polymorphisms, allowing fragment size detection and analysis by automated capillary electrophoresis or similar techniques. The species identification is achieved by comparing the target sample profile (DNA fragment sizes) with pre-established standard species-specific fragment sizes. With the present invention is possible to detect and identify animal species in highly degraded biological samples due to the analysis of mitochondrial DNA, present in many copies per cell. This invention is advantageous when high throughput sample analyses are needed such as for large-scale species identification procedures in forensics, food quality control, ancient DNA, paleogenetics and ecology studies. Another aspect of the present invention relates to a kit containing reagents needed to practice the above-referred method.
机译:本发明涉及基于线粒体12s和16s核糖体RNA基因序列的大小的变化,在具有动物来源遗传物质的生物样品中鉴定动物种类的方法。本发明的方法包括一组用荧光染料标记的寡核苷酸PCR(聚合酶链反应)引物,用于多重PCR扩增具有插入/缺失多态性的七个线粒体DNA区域,从而可以通过自动毛细管电泳或类似方法检测片段大小并进行分析。技术。物种鉴定是通过将​​目标样品图谱(DNA片段大小)与预先建立的标准物种特异性片段大小进行比较来实现的。利用本发明,由于线粒体DNA的分析,可以检测和鉴定高度降解的生物样品中的动物种类,线粒体DNA每个细胞以许多拷贝存在。当需要高通量样品分析时,例如在法医,食品质量控制,古代DNA,古生物学和生态学研究中进行大规模物种鉴定程序时,本发明是有利的。本发明的另一方面涉及一种试剂盒,其包含实施上述方法所需的试剂。

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