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Laser microdissection and DNA typing of cells from single hair follicles.

机译:激光显微切割和单毛囊细胞的DNA分型。

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摘要

Isolation and identification of single cells from tissue samples or smears assume a great relevance in pathological and forensic applications; in this latter field, the possibility to identify a specific genetic profile can be obtained by short tandem repeat (STR) typing, allowing to achieve a scientific proof important in law courts. It is well known that DNA extraction may be performed from several tissue fragments, blood traces, spermatozoa as well as telogen hair. However, in the last case, few follicle cells are coupled to a great amount of keratin reducing the efficiency of DNA amplification. Recently, the introduction of laser microdissection technique has greatly improved the capability to select single cells without any cross-contamination. In the present report, we have performed a laser microdissection using a Leica AS LMD (Leica Microsystems, Germany), utilized on cutting the telogen hair in order to exclusively collect the lower part of the follicle and reduce keratin contamination. In this way we can accurately extract an adequate amount of DNA, successfully typed by STR profile.
机译:从组织样本或涂片中分离和鉴定单个细胞在病理学和法医应用中具有重要的意义。在后一个领域中,可以通过短串联重复序列(STR)打字获得鉴定特定遗传特征的可能性,从而获得在法庭上重要的科学证据。众所周知,可以从几个组织片段,血液痕迹,精子以及毛发生长的头发中提取DNA。但是,在最后一种情况下,很少有卵泡细胞与大量的角蛋白偶联,从而降低了DNA扩增的效率。最近,激光显微切割技术的引入大大提高了选择单个细胞而没有任何交叉污染的能力。在本报告中,我们使用Leica AS LMD(德国Leica Microsystems)进行了激光显微切割,该激光显微切割用于切割毛发期末期毛发,以便专门收集卵泡下部并减少角蛋白污染。通过这种方式,我们可以准确地提取足够数量的DNA,并通过STR谱图成功地进行了键入。

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