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Genotyping genetic markers from LCN and degraded DNA by HRM and their application in hair shaft

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Abstract

Degraded and low copy number (LCN) DNA samples are common challenging materials in forensic casework because they increase the difficulty of sample processing and reduce the possibility of obtaining genetic information from DNA. High-resolution melting (HRM) curve analysis is promising for genotyping genetic markers and has been applied to the detection of LCN and degraded DNA in the field of forensic science. However, the exact assessment based on HRM at multiple genetic markers from both degraded and LCN DNA materials has not been optimized. To explore the ability of HRM to genotype LCN and degraded DNA samples, we selected three genetic markers to genotype in experimental LCN and degraded DNA and practical hair shaft materials, which are often encountered as degraded and LCN DNA in forensic medicine. The results show that DNA samples of as low as 100 pg and as short as 60 bp were successfully genotyped by the HRM assay at all three genetic markers, whereas in hair shaft DNA, two loci were accurately genotyped. The HRM assay established in this study can be applied to LCN and degraded DNA analysis in forensic casework and can act as a reference point before genotyping short tandem repeat markers. Developing the HRM strategy for genotyping DNA genetic markers enriches detectable targets in hair shaft samples and provides valuable data for further exploration.

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Funding

The present study was supported by grant from the National Natural Science Foundation of China (grant no. 81471826).

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Authors and Affiliations

  1. Department of Forensic Genetics and Biology, School of Forensic Medicine, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang, 110122, People’s Republic of China

    Enzhu Jiang, Siyi Zhang & Hao Pang

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  1. Enzhu Jiang
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  2. Siyi Zhang
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  3. Hao Pang
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Correspondence to Hao Pang.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Fig. S1

PCR efficiencies of c.261delG (a) and c.803G>C (b) in the ABO gene and a 3-bp insertion in the AMELY gene (c) (PNG 174 kb)

High Resolution Image (TIF 1482 kb)

Fig. S2

Genotyping result of a 3-bp insertion with mixed X- and Y-chromosome-specific reverse primers, using a gDNA template (a), LCN template (b), or degraded template (c). The PCR efficiencies are given (d) (PNG 318 kb)

High Resolution Image (TIF 2122 kb)

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Jiang, E., Zhang, S. & Pang, H. Genotyping genetic markers from LCN and degraded DNA by HRM and their application in hair shaft. Int J Legal Med 134, 31–37 (2020). https://doi.org/10.1007/s00414-019-02045-w

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  • DOI: https://doi.org/10.1007/s00414-019-02045-w

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