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Pyrosequencing of a short fragment of the amelogenin gene for gender identification

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Abstract

We report a pyrosequencing method for detecting a short amelogenin fragment to aid the gender identification. The PCR products (44/45 bp), including primers and target sequence (4/5 bp) consisting of three point mutations and one indel mutation, were sequenced by the pyrosequencing method. 100 randomly chosen DNA samples of healthy donors were analyzed with this method, and all of them were correctly typed. The sensitivity of the technique was 0.5 ng template DNA. No specific peak was found in any detected animals or organisms except for monkey. For blood samples that were left outside for 26 weeks and DNA degraded artificially by digesting with DNaseI, this method gave more accurate results than the conventional method. Moreover, four bone samples analyzed using the method gave clear pyrograph. This method is easy, quick, cheap and suitable for high-throughput analysis, especially for identifying the gender of highly-degraded DNA samples.

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Abbreviations

PAGE:

Polyacrylamide gel electrophoresis

PCR:

Polymerase chain reaction

SNP:

Single nucleotide polymorphism

STR:

Short tandem repeats

AMEL:

Amelogenin

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Acknowledgments

We thank Lei Zhao, Xue Bai for their support on bone DNA extraction techniques, and Bangxian Dou, Jixing Zhu for their support on Bone samples supply. This study was supported by National Natural Science Foundation of China (30973364) and Natural Science Foundation of Hebei Province (C2010000459).

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Authors and Affiliations

  1. Hebei Key Laboratory of Forensic Medicine, Department of Forensic Medicine, Hebei Medical University, No. 361 Zhong Shan Road, Shijiazhuang, 050017, Hebei, People’s Republic of China

    Shujin Li, Ting Feng, Lihong Fu, Chunguang Lou, Xiaojing Zhang, Chunling Ma & Bin Cong

  2. Department of Thoracic Surgery, The Fourth Affiliated Hospital, Hebei Medical University, No. 12 Jian Kang Road, Shijiazhuang, 050011, Hebei, People’s Republic of China

    Zhenhua Li

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  1. Shujin Li
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  2. Ting Feng
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  3. Lihong Fu
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  4. Zhenhua Li
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  5. Chunguang Lou
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  6. Xiaojing Zhang
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  7. Chunling Ma
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  8. Bin Cong
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Corresponding author

Correspondence to Bin Cong.

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Shujin Li and Ting Feng contributed equally to this work.

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Li, S., Feng, T., Fu, L. et al. Pyrosequencing of a short fragment of the amelogenin gene for gender identification. Mol Biol Rep 39, 6949–6957 (2012). https://doi.org/10.1007/s11033-012-1522-2

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  • DOI: https://doi.org/10.1007/s11033-012-1522-2

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