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Assessing a novel room temperature DNA storage medium for forensic biological samples

机译:评估用于法医生物样品的新型室温DNA存储介质

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The ability to properly collect, analyze and preserve biological stains is important to preserving the integrity of forensic evidence. Stabilization of intact biological evidence in cells and the DNA extracts from them is particularly important since testing is generally not performed immediately following collection. Furthermore, retesting of stored DNA samples may be needed in casework for replicate testing, confirmation of results, and to accommodate future testing with new technologies. A novel room temperature DNA storage medium, SampleMatrix? (SM; Biomatrica, Inc., San Diego, CA), was evaluated for stabilizing and protecting samples. Human genomic DNA samples at varying amounts (0.0625-200 ng) were stored dry in SM for 1 day to 1 year under varying conditions that included a typical ambient laboratory environment and also through successive freeze-thaw cycles (3 cycles). In addition, spiking of 1-4× SM into samples prior to analysis was performed to determine any inhibitory effects of SM. Quantification of recovered DNA following storage was determined by quantitative PCR or by agarose gel electrophoresis, and evaluation of quantitative peak height results from multiplex short tandem repeat (STR) analyses were performed to assess the efficacy of SM for preserving DNA. Results indicate no substantial differences between the quality of samples stored frozen in liquid and those samples maintained dry at ambient temperatures protected in SM. For long-term storage and the storage of low concentration samples, SM provided a significant advantage over freezer storage through higher DNA recovery. No detectable inhibition of amplification was observed at the recommended SM concentration and complete profiles were obtained from genomic DNA samples even in the presence of higher than recommended concentrations of the SM storage medium. The ability to stabilize and protect DNA from degradation at ambient temperatures for extended time periods could have tremendous impact in simplifying and improving sample storage conditions and requirements. The current work focuses on forensics analysis; however this technology is applicable to all endeavors requiring storage of DNA.
机译:正确收集,分析和保存生物污渍的能力对于保持法医证据的完整性很重要。稳定细胞中完整的生物学证据以及从其中提取DNA的方法尤为重要,因为通常不会在收集后立即进行检测。此外,在个案工作中可能需要对存储的DNA样本进行重新测试,以进行重复测试,结果确认以及适应将来使用新技术进行的测试。一种新型的室温DNA存储介质SampleMatrix? (SM; Biomatrica,Inc。,圣地亚哥,CA)被评估用于稳定和保护样品。将不同量(0.0625-200 ng)的人类基因组DNA样品在不同条件下(包括典型的环境实验室环境)并通过连续的冻融循环(3个循环)在SM中干燥保存1天至1年。另外,在分析之前将1-4x SM掺入样品中以确定SM的任何抑制作用。通过定量PCR或琼脂糖凝胶电泳确定存储后回收的DNA的定量,并通过多重短串联重复序列(STR)分析对定量峰高结果进行评估,以评估SM用于保存DNA的功效。结果表明,冷冻保存在液体中的样品质量与那些在SM中保护的环境温度下保持干燥的样品之间没有实质性差异。对于长期存储和低浓度样品的存储,SM通过更高的DNA回收率提供了优于冷冻存储的显着优势。在推荐的SM浓度下,未观察到可检测到的扩增抑制作用,即使存在高于推荐浓度的SM存储介质,也可从基因组DNA样品中获得完整的图谱。稳定和保护DNA在环境温度下长时间不降解的能力可能对简化和改善样品存储条件和要求产生巨大影响。目前的工作重点是取证分析。但是,该技术适用于所有需要存储DNA的工作。

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