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用于检测火星土壤模拟物中最低浓度DNA的灵敏方法的开发。

Development of Sensitive Methods for the Detection of Minimum Concentrations of DNA on Martian Soil Simulants.

作者信息

Li Yongda, Rochfort Keith D, Collins David, Grintzalis Konstantinos

机构信息

School of Biotechnology, Dublin City University, D09 Y5NO Dublin, Ireland.

School of Nursing, Psychotherapy, and Community Health, Dublin City University, D09 Y5NO Dublin, Ireland.

出版信息

Life (Basel). 2023 Sep 30;13(10):1999. doi: 10.3390/life13101999.

Abstract

Several methods used for the quantification of DNA are based on UV absorbance or the fluorescence of complexes with intercalator dyes. Most of these intercalators are used in gels to visualize DNA and its structural integrity. Due to many extraterrestrial samples, such as meteorites or comets, which are likely to contain very small amounts of biological material, and because the ability to detect this material is crucial for understanding the origin and evolution of life in the universe, the development of assays that can detect DNA at low limits and withstand the rigors of space exploration is a pressing need in the field of astrobiology. In this study, we present a comparison of optimized protocols used for the fast and accurate quantification of DNA using common intercalator dyes. The sensitivity of assays exceeded that generated by any commercial kit and allowed for the accurate quantification of minimum concentrations of DNA. The methods were successful when applied to the detection and measurement of DNA spiked on soil samples. Furthermore, the impact of UV radiation as a harsh condition on the surface of Mars was assessed by DNA degradation and this was also confirmed by gel electrophoresis. Overall, the methods described provide economical, simple-step, and efficient approaches for the detection of DNA and can be used in future planetary exploration missions as tests used for the extraction of nucleic acid biosignatures.

摘要

几种用于DNA定量的方法是基于紫外线吸光度或与嵌入染料形成的复合物的荧光。这些嵌入剂大多用于凝胶中以观察DNA及其结构完整性。由于许多外星样本,如陨石或彗星,可能含有极少量的生物物质,而且检测这种物质的能力对于理解宇宙中生命的起源和演化至关重要,因此开发能够在低限度检测DNA并经受住太空探索严苛条件的检测方法是天体生物学领域的迫切需求。在本研究中,我们比较了使用常见嵌入染料快速准确地定量DNA的优化方案。这些检测方法的灵敏度超过了任何商业试剂盒所产生的灵敏度,并能够准确地定量最低浓度的DNA。这些方法应用于检测和测量添加到土壤样本中的DNA时取得了成功。此外,通过DNA降解评估了紫外线辐射作为火星表面恶劣条件的影响,这也通过凝胶电泳得到了证实。总体而言,所描述的方法为DNA检测提供了经济、简单且高效的途径,可用于未来的行星探索任务,作为用于提取核酸生物特征的测试方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1dc2/10608431/58181d0b0963/life-13-01999-g001.jpg

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