精子富集柱分离技术的研究
发布时间：2018-08-24 15:56
【摘要】：目的在传统差异裂解法基础上,研究建立新的混合斑检材中精子分离技术。方法根据精细胞的结构特点,研制精子富集柱。取混合斑检材经第一步消化后的混合液,加入精子富集柱中离心,使DNA等小分子物质透过,而精细胞被特异性黏附和拦截在富集层中。采用本文技术和常规裂解方法对12份混合斑检材中精子进行分离,分离后精子DNA采用Identifiler试剂盒进行PCR扩增和电泳检测。结果混合斑检材经精子富集柱分离后均获得单一男性精子的STR分型结果,而12份检材用常规裂解方法分离,其中有6份检材女性成分去除不完全或未能检出精子STR基因型。结论本研究建立的精子富集柱分离技术适用于常见混合斑检材中精子的分离,特别适合对含有大量女性混合物而精子量较少检材的分离提取。
[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.
【作者单位】： 辽宁省刑事科学技术研究所;辽宁大学生命科学院;
【分类号】：D919
[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.
【作者单位】： 辽宁省刑事科学技术研究所;辽宁大学生命科学院;
【分类号】：D919
【相似文献】
相关期刊论文 前10条
1 封宇;刘云;吕晓革;修崇贤;;混合斑中精子细胞分离实验条件探讨[J];中国法医学杂志;2009年01期
2 袁自闯;赵凯;李海燕;孙宏钰;欧桂生;;Differex~(TM)系统在混合斑精子DNA分离提取中的应用[J];中国法医学杂志;2006年02期
3 吴洪槟;;混合斑的DNA鉴定的几种方法研究[J];攀枝花学院学报;2007年06期
4 董研;陈荣华;平原;郑会芬;;应用Differex~(TM)法提取混合斑中精子DNA[J];中国法医学杂志;2006年05期
5 焦德利;孙树毅;李海波;;成功检验保存10年的精斑1例[J];刑事技术;2011年03期
6 李鑫;胡兰;郭红玲;刘志芳;Q谜，
本文编号：2201307
本文链接：https://www.wllwen.com/falvlunwen/fanzuizhian/2201307.html
