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

被引：1

作者：

Harikai, Naoki ^{[1
]}

Takada, Yuki ^{[1
]}

Saito, Misaki ^{[1
]}

Zaima, Kazumasa ^{[1
]}

Shinomiya, Kazufusa ^{[1
]}

机构：

[1] Nihon Univ, Sch Pharm, 7-7-1 Narashinodai, Funabashi 2748555, Japan

来源：

BIOPRESERVATION AND BIOBANKING | 2024年 / 22卷 / 03期

关键词：

autoclave; DNA damage; DNA degradation; DNA fragmentation; PCR; thermal degradation; REAL-TIME PCR; QUANTITATIVE PCR; QUANTIFICATION; DEGRADATION; FRAGMENTATION; DAMAGE; MAIZE;

D O I：

10.1089/bio.2023.0021

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

This study examined the influence of heat exposure on DNA samples during polymerase chain reaction (PCR) detection. In this study, lambda DNA samples, as model DNA, were exposed to 105 degrees C for 3-90 minutes or to 105 degrees C-115 degrees 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 degrees C by autoclaving, the data showed negative correlations between the logarithm of lambda DNA concentration (log lambda DNA) calculated using real-time PCR and exposure duration and a good relationship between the slope of the regression line and amplicon size. Regarding lambda DNA samples exposed to heat for 15 minutes, the data showed negative correlations between the log lambda DNA 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 lambda DNA samples by autoclaving, and the PCR detection levels of the DNA at each amplicon size.

引用

页码：268 / 274

页数：7

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