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Mismatched Multiplex PCR Amplification and Subsequent RFLP Analysis to Simultaneously Identify Polymorphisms of Erythrocytic ESD, GLO1, and GPT Genes

Authors

  • Hao Pang Ph.D.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Ye Ding B.S.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Yan Li B.S.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Lizhi Wang B.S.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Xiaofei Tian M.S.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Baojie Wang Ph.D.,

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Mei Ding Ph.D.

    1. Department of Forensic Serology, School of Forensic Medicine, China Medical University, 92 Beier Road, Heping District, Shenyang 110001, China.
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  • Supported by a grant from the Educational and Scientific Foundation of Liaoning Provincial Education Department of China and initial funding from China Medical University.

Additional information and reprint requests:
Hao Pang, Ph.D.
Professor
Department of Forensic Serology
China Medical University
92 Beier Road, Heping District
Shenyang 110001
China
E-mail: panghao8@gmail.com

Abstract

Abstract: ESD (esterase D), GLO1 (glyoxalase I), and GPT (glutamate pyruvate transaminase) are human erythrocytic isoenzymes and have previously been applied in forensic medicine caseworks. The molecular bases of the polymorphic gene expression products have been demonstrated to be because of SNPs in respective coding regions. However, it has not been revealed whether the SNPs conferring the polymorphisms to the aforementioned erythrocytic isoenzymes could be simultaneously detected by using a simple PCR method. In this study, we used mismatched primers to simultaneously amplify three common isoenzyme loci so that all amplified products contained the same Hph I cleavage sites. The products were then digested with Hph I and electrophoretically separated and stained so that alleles were identified. The accumulated values for the probability of discrimination power and excluding the probability of paternity to the aforementioned systems attained 90.41% and 41.72%, respectively, in the Chinese Han population. This assay could be extremely valuable for future forensic medicine practices.

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