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优化DNA提取物的储存与处理

Optimizing Storage and Handling of DNA Extracts.

作者信息

Lee S B, Crouse C A, Kline M C

机构信息

Forensic Science Program, Justice Studies Department, San Jose State University, San Jose, CA, USA.

Forensic Biology Unit, Palm Beach County Sheriff's Office Crime Laboratory, West Palm Beach, FL, USA.

出版信息

Forensic Sci Rev. 2010 Jul;22(2):131-44.

Abstract

Nucleic acid sample storage is of paramount importance in forensic science as well as in epidemiological, clinical, and genetic laboratories. Millions of biological samples, including cells, viruses, and DNA/RNA, are stored every year for diagnostics, research, and forensic science. PCR has permitted the analysis of minute sample quantities. Samples such as bone, teeth, touch samples, and some sexual assault evidence may yield only low-quality and low-quantity DNA/RNA. Efficient storage of the extracted DNA/RNA is needed to ensure the stability of the sample over time for retesting of the CODIS STRs, mtDNA, YSTRs, mRNA, and other future marker-typing systems. Amplification of some or all of these markers may fail because the biological material has been highly degraded, contains inhibitors, is too low in quantity, or is contaminated with contemporary DNA. Reduction in recovery has been observed with refrigerated liquid DNA extracts and also those exposed to multiple freeze-thaw cycles. Therefore, the development of optimal storage and amplification methods is critical for successful recovery of profiles from these types of samples since, in many cases, retesting is necessary. This review is divided into three sections. The Introduction and Background covers forensic DNA storage, factors that influence DNA stability, and a brief review of molecular strategies to type non-optimal DNA. Section I covers the importance of DNA extract storage in forensic and non-forensic DNA databanks and the mechanisms responsible for loss during storage. Finally, Section II covers strategies and technologies being utilized to store DNA.

摘要

核酸样本储存在法医学以及流行病学、临床和基因实验室中至关重要。每年都有数以百万计的生物样本被储存,包括细胞、病毒以及DNA/RNA,用于诊断、研究和法医学。聚合酶链反应(PCR)使得对微量样本的分析成为可能。诸如骨骼、牙齿、接触样本以及一些性侵犯证据等样本可能只能产生低质量和低数量的DNA/RNA。为确保样本随时间推移的稳定性以便对联合DNA索引系统(CODIS)的短串联重复序列(STR)、线粒体DNA(mtDNA)、Y染色体STR(YSTR)、信使核糖核酸(mRNA)以及其他未来标记分型系统进行重新检测,需要对提取的DNA/RNA进行有效储存。由于生物材料已高度降解、含有抑制剂、数量过低或被当代DNA污染,对这些标记中的部分或全部进行扩增可能会失败。已观察到冷藏的液体DNA提取物以及经历多次冻融循环的提取物的回收率会降低。因此,开发最佳的储存和扩增方法对于从这类样本中成功恢复图谱至关重要,因为在许多情况下有必要进行重新检测。本综述分为三个部分。引言和背景部分涵盖法医DNA储存、影响DNA稳定性的因素以及对非最佳DNA分型的分子策略的简要回顾。第一部分涵盖DNA提取物储存在法医和非法医DNA数据库中的重要性以及储存期间损失的原因。最后,第二部分涵盖用于储存DNA的策略和技术。

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