Crouse C A, Ban J D, D'Alessio J K
Crime Laboratory, Palm Beach County Sheriff's Office, West Palm Beach, FL 33406.
Biotechniques. 1993 Oct;15(4):641-4, 646, 648.
Sonication procedures for the extraction of DNA from forensic-type semen specimens have been developed, which, when compared to currently utilized sperm DNA extraction techniques, are simple, rapid and result in comparable DNA yields. Sperm DNA extraction by sonication was performed on whole semen, seminal stains, buccal swabs and post-coital specimens. Ultrasound disruption of sperm cells and their ultimate release of cellular DNA has been conducted in the presence of sperm wash buffers followed by organic extraction or Chelex 100 with little or no compromise to DNA quality, quantity or amplifiability. Two advantages of sonication over currently used forensic techniques to extract sperm DNA include 1) sperm DNA extraction that occurs within five minutes of sonication compared with an hour or greater for water bath incubations in classic enzyme digestion DNA extractions and 2) one less preparatory step with the Chelex/sonication protocol and three less steps with the sonication/organic protocol compared with other procedures thus eliminating potential sample-to-sample cross-contamination. Sperm DNA extracted by optimum sonication procedures was used for forensic HLA DQ alpha typing and restriction fragment length polymorphisms analysis without any adverse effects on typing results.
已开发出用于从法医类型精液样本中提取DNA的超声处理程序，与目前使用的精子DNA提取技术相比，该程序简单、快速，且DNA产量相当。通过超声处理从全精液、精液污渍、口腔拭子和性交后样本中提取精子DNA。在精子洗涤缓冲液存在的情况下，对精子细胞进行超声破碎及其细胞DNA的最终释放，随后进行有机提取或使用Chelex 100，对DNA质量、数量或可扩增性几乎没有损害。超声处理相对于目前用于提取精子DNA的法医技术的两个优点包括：1）超声处理后五分钟内即可完成精子DNA提取，而经典酶消化DNA提取中的水浴孵育则需要一小时或更长时间；2）与其他程序相比，Chelex/超声处理方案少一个准备步骤，超声处理/有机方案少三个步骤，从而消除了潜在的样本间交叉污染。通过最佳超声处理程序提取的精子DNA用于法医HLA DQα分型和限制性片段长度多态性分析，对分型结果没有任何不利影响。
