Raman Spectroscopy: A Promising Technique for Analyzing Nucleic Acid Refractory to PCR Amplification

Milad Soleimani, Rachel Matar, Maxime Merheb
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引用次数: 2

Abstract

Ancient DNA (aDNA) refers to the genetic material found in dead paleontological and archeological samples. Being subject to various types of stressors, it undergoes different hydrolytic and oxidative post mortem modifications that result in the formation of DNA lesions. These lesions are found to either block the DNA polymerase during replication or induce nucleotide misincorporations. Besides, aDNA samples occur in minimal quantities; which represents an additional obstacle that researchers have to overcome in order to study aDNA. The earliest major efforts included the use of molecular cloning and polymerase chain reaction (PCR) to amplify aDNA sequences. These techniques were later found to be associated with a number of false results and unauthentic findings. There have been numerous attempts to eliminate the shortcomings of PCR and improve the quality of aDNA through avoiding contamination, repairing lesions, using translesion polymerases, etc. However, the majority of these have failed to yield accurate and specific results. Surface- enhanced resonance Raman scattering (SERRS), on the other hand, starts in the right foot by introducing a sensitive non- enzymatic approach for the specific detection of single- and double-stranded DNA. The ability of this method to evade DNA degradation is particularly important for not only studying aDNA, but also analyzing DNA refractory to PCR amplification in processed products.
拉曼光谱:一种有前途的分析难以PCR扩增的核酸的技术
古DNA (aDNA)是指在古生物和考古死亡样本中发现的遗传物质。由于受到各种应激源的影响,它在死后会发生不同的水解和氧化修饰,从而导致DNA损伤的形成。这些病变被发现在复制过程中阻断DNA聚合酶或诱导核苷酸错结合。此外,aDNA样本的数量很少;这是研究人员为了研究aDNA必须克服的另一个障碍。最早的主要努力包括使用分子克隆和聚合酶链反应(PCR)来扩增aDNA序列。这些技术后来被发现与一些错误的结果和不真实的发现有关。人们曾多次尝试通过避免污染、修复病变、使用翻译聚合酶等方法来消除PCR的缺点,提高aDNA的质量。然而,其中大多数都未能产生准确和具体的结果。另一方面,表面增强共振拉曼散射(SERRS)从右脚开始,引入了一种敏感的非酶方法,用于特异性检测单链和双链DNA。这种方法逃避DNA降解的能力不仅对研究aDNA,而且对分析加工产物中难以PCR扩增的DNA尤为重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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