【摘要】：目的在傳統(tǒng)差異裂解法基礎(chǔ)上,研究建立新的混合斑檢材中精子分離技術(shù)。方法根據(jù)精細(xì)胞的結(jié)構(gòu)特點(diǎn),研制精子富集柱。取混合斑檢材經(jīng)第一步消化后的混合液,加入精子富集柱中離心,使DNA等小分子物質(zhì)透過,而精細(xì)胞被特異性黏附和攔截在富集層中。采用本文技術(shù)和常規(guī)裂解方法對(duì)12份混合斑檢材中精子進(jìn)行分離,分離后精子DNA采用Identifiler試劑盒進(jìn)行PCR擴(kuò)增和電泳檢測(cè)。結(jié)果混合斑檢材經(jīng)精子富集柱分離后均獲得單一男性精子的STR分型結(jié)果,而12份檢材用常規(guī)裂解方法分離,其中有6份檢材女性成分去除不完全或未能檢出精子STR基因型。結(jié)論本研究建立的精子富集柱分離技術(shù)適用于常見混合斑檢材中精子的分離,特別適合對(duì)含有大量女性混合物而精子量較少檢材的分離提取。
[Abstract]:Objective to establish a new technique for sperm separation from mixed spot samples based on traditional differential cleavage method. Methods according to the structural characteristics of spermatocytes, spermatozoa enrichment column was prepared. After the mixed spot samples were digested by the first step, the mixture was added to the sperm enrichment column to centrifuge, so that DNA and other small molecular substances were passed through, while the spermatocytes were specifically adhered and intercepted in the enrichment layer. The spermatozoa from 12 mixed spot samples were separated by using the technique of routine cleavage and the PCR amplification and electrophoretic detection of sperm DNA were carried out by Identifiler kit. Results the STR typing results of single male spermatozoa were obtained after the mixed spot samples were separated by spermatozoa enrichment column, while 12 samples were separated by routine cleavage method. Among them, 6 samples were female components removed incomplete or failed to detect sperm STR genotypes. Conclusion the spermatozoa enrichment column technique established in this study is suitable for the separation of spermatozoa from common mixed spot samples, especially for the separation and extraction of spermatozoa containing a large amount of female mixture but less sperm quantity.
【作者單位】： 遼寧省刑事科學(xué)技術(shù)研究所;遼寧大學(xué)生命科學(xué)院;
【分類號(hào)】：D919

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本文編號(hào)：2201307