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采用 TaqMan 分析技术评估和 SNP 分型经紫外线照射后的人血斑中的 DNA。

Evaluation and SNP typing of DNA from ultraviolet-irradiated human bloodstains using TaqMan assay.

机构信息

Division of Legal Medicine, Department of Social Medicine, Nihon University School of Medicine, Tokyo, 1738610, Japan.

出版信息

Sci Rep. 2021 Apr 13;11(1):8029. doi: 10.1038/s41598-021-87313-9.

Abstract

When detecting DNA profiles from forensic materials, it is pivotal to know the extent of degradation and which DNA marker can be genotyped. Ultraviolet (UV) is one of the common external factors that causes DNA damage, through which, an attempt to reveal cardinal genetic information can be made. In this study, after irradiation with three different UV wavelengths, UV-damaged DNA in the bloodstains was analyzed with long and short TaqMan assays using real-time PCR. In addition, both short tandem repeat (STR) profiles and single nucleotide polymorphisms (SNPs) from the damaged DNA at different stages of UV exposure were also assessed. With increasing in UV irradiation cycles, there was a delay of the amplification curves accompanied with a decrease in the DNA amounts collected. Despite the amplification of STR genotype was not altered after 75 cycles of UVC irradiation, all 12 SNP loci could still be detected. Furthermore, a short-assay line was detected in the absence of an amplification of the evaluation curve. The results indicate that, although the DNA template might not be useful and suitable for analysis of STR profile, this approach is of some values in detecting SNPs.

摘要

在法医材料的 DNA 图谱检测中,了解降解程度以及哪些 DNA 标记可以进行基因分型至关重要。紫外线 (UV) 是导致 DNA 损伤的常见外部因素之一,通过它可以尝试揭示重要的遗传信息。在这项研究中,在三种不同的 UV 波长照射后,使用实时 PCR 分析了血液斑中经紫外线损伤的 DNA 的长和短 TaqMan 检测。此外,还评估了在不同紫外线照射阶段受损 DNA 的短串联重复 (STR) 图谱和单核苷酸多态性 (SNP)。随着 UV 照射循环次数的增加,扩增曲线出现延迟,同时收集的 DNA 量减少。尽管在 UVC 照射 75 个循环后,STR 基因型的扩增没有改变,但仍可以检测到 12 个 SNP 位点。此外,在评估曲线没有扩增的情况下,检测到了短检测线。结果表明,尽管 DNA 模板可能不适用于 STR 图谱的分析,但这种方法在检测 SNP 方面具有一定的价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935c/8044161/9dd643a3d774/41598_2021_87313_Fig1_HTML.jpg

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