当前位置: X-MOL 学术J. Mammal. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Temporal-dependent effects of DNA degradation on frozen tissues archived at −80°C
Journal of Mammalogy ( IF 1.5 ) Pub Date : 2021-02-03 , DOI: 10.1093/jmammal/gyab009
Taylor J Soniat 1 , Hendra F Sihaloho 1 , Richard D Stevens 2, 3 , Todd D Little 4 , Caleb D Phillips 1, 2 , Robert D Bradley 1, 2
Affiliation  

Frozen tissues, associated with natural history and biological collections, historically have been archived at temperatures between −20°C and −80°C. More recently, the availability of liquid nitrogen systems has enabled the storage of tissue samples (biobanking) at temperatures as low as −196°C. Currently, it is not known how the degree of coldness (e.g., −80°C or −196°C) or longevity (time in storage) impacts preservation of tissue samples. To examine the effects of long-term storage (−80°C and −196°C) on DNA degradation, tissue samples (muscle and liver) archived for 30, 20, 10, or 1 years were obtained from the Natural Science Research Laboratory at Texas Tech University. The integrity of DNA (measured as molecular weight and fragment length) extracted from samples was determined using automated DNA isolation methods followed by microfluidic distribution measurement. DNA distributions were compared using measures of central tendency, a regression-based molecular mass profile, and as a latent variable in a structural equation model. Muscle samples consistently outperformed liver samples in terms of quality of DNA yield. Also, muscle samples exhibited a significant linear relationship with time in which older samples were more degraded than were recent samples. The signal for a temporal effect on DNA was strongest when considering a latent variable of DNA quality based on mode and kurtosis; 37% of the variation in the latent variable was explained by variation in units of time. More recent time points tended to be more similar, but the temporal effect on the latent variable remained strong even when the oldest samples were removed from the analysis. In contrast, integrity of DNA from liver samples did not have a significant linear relationship with time; however, in some years they exhibited non-normally distributed DNA quality metrics that may have reflected sensitivity of liver tissue to degradation during specimen preparation, DNA extraction, or archive parameters. Results indicated that tissue type and temporal effects influenced rates of DNA degradation, with the latter emphasizing the long-term value of biobanking at the coldest temperatures possible (liquid nitrogen storage) to mitigate degradation of biological samples of ever-increasing scientific value.

中文翻译:

DNA降解对-80°C冷冻组织的时间依赖性影响

与自然史和生物收藏相关的冷冻组织历来在 -20°C 和 -80°C 之间的温度下存档。最近,液氮系统的可用性使得组织样本(生物库)能够在低至 -196°C 的温度下储存。目前,尚不清楚寒冷程度(例如,-80°C 或-196°C)或寿命(储存时间)如何影响组织样本的保存。为了检查长期储存(-80°C 和 -196°C)对 DNA 降解的影响,从自然科学研究实验室获得了 30、20、10 或 1 年存档的组织样本(肌肉和肝脏)在德克萨斯理工大学。使用自动 DNA 分离方法确定从样品中提取的 DNA 的完整性(测量为分子量和片段长度),然后进行微流体分布测量。DNA 分布使用集中趋势测量、基于回归的分子量分布和作为结构方程模型中的潜在变量进行比较。就 DNA 产量的质量而言,肌肉样本始终优于肝脏样本。此外,肌肉样本与时间呈显着的线性关系,其中较旧的样本比最近的样本退化得更多。当考虑基于众数和峰度的 DNA 质量潜在变量时,对 DNA 的时间影响的信号最强;潜变量中 37% 的变化可以用时间单位的变化来解释。最近的时间点往往更相似,但即使从分析中删除最旧的样本,对潜在变量的时间影响仍然很强。相比之下,来自肝脏样本的 DNA 完整性与时间没有显着的线性关系。然而,在某些年份,它们表现出非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在标本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。但是即使从分析中删除最旧的样本,对潜在变量的时间影响仍然很强。相比之下,来自肝脏样本的 DNA 完整性与时间没有显着的线性关系。然而,在某些年份,它们表现出非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在标本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。但是即使从分析中删除最旧的样本,对潜在变量的时间影响仍然很强。相比之下,来自肝脏样本的 DNA 完整性与时间没有显着的线性关系。然而,在某些年份,它们表现出非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在标本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。来自肝脏样本的 DNA 完整性与时间没有显着的线性关系;然而,在某些年份,它们表现出非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在标本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。来自肝脏样本的 DNA 完整性与时间没有显着的线性关系;然而,在某些年份,它们表现出非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在标本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。在某些年份,他们展示了非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在样本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。在某些年份,他们展示了非正态分布的 DNA 质量指标,这些指标可能反映了肝组织在样本制备、DNA 提取或存档参数期间对降解的敏感性。结果表明,组织类型和时间效应影响 DNA 降解率,后者强调了生物库在可能的最冷温度(液氮储存)下的长期价值,以减轻具有不断增加的科学价值的生物样品的降解。
更新日期:2021-02-03
down
wechat
bug