Relationship Between Amplicon Size and Heat Conditions in Polymerase Chain Reaction Detection of DNA Degraded by Autoclaving.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-06-01 Epub Date: 2023-10-23 DOI:10.1089/bio.2023.0021

Naoki Harikai, Yuki Takada, Misaki Saito, Kazumasa Zaima, Kazufusa Shinomiya

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Abstract

This study examined the influence of heat exposure on DNA samples during polymerase chain reaction (PCR) detection. In this study, λDNA samples, as model DNA, were exposed to 105°C for 3-90 minutes or to 105°C-115°C for 15 minutes by autoclaving. The exposed samples were subjected to real-time PCR using nine primer sets with amplicon sizes of 45-504 bp. Regarding DNA samples exposed to 105°C by autoclaving, the data showed negative correlations between the logarithm of λDNA concentration (log λDNA) calculated using real-time PCR and exposure duration and a good relationship between the slope of the regression line and amplicon size. Regarding λDNA samples exposed to heat for 15 minutes, the data showed negative correlations between the log λDNA and exposure temperature and a good relationship between the slope of the regression line and amplicon size. These results showed that the equations used in this study could predict the degree of degradation in λDNA samples by autoclaving, and the PCR detection levels of the DNA at each amplicon size.

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聚合酶链反应检测高压灭菌降解DNA中扩增子大小与加热条件的关系。

本研究检测了聚合酶链式反应（PCR）检测过程中热暴露对DNA样本的影响。在本研究中，λDNA样品作为模型DNA，通过高压灭菌暴露于105°C下3-90分钟或105°C-115°C下15分钟。使用扩增子大小为45-504的九个引物组对暴露的样品进行实时PCR bp。关于通过高压灭菌暴露于105°C的DNA样本，数据显示，使用实时PCR计算的λDNA浓度的对数（logλDNA）与暴露时间呈负相关，回归线的斜率与扩增子大小之间存在良好关系。关于暴露于高温15分钟的λDNA样本，数据显示logλDNA与暴露温度之间呈负相关，回归线的斜率与扩增子大小之间呈良好关系。这些结果表明，本研究中使用的方程可以预测高压灭菌后λDNA样品的降解程度，以及每个扩增子大小下DNA的PCR检测水平。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.

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