alexa Forensic DNA Typing of Old Skeletal Remains Using AmpFlSTR® Identifiler® PCR Amplification Kit
ISSN: 2157-7145

Journal of Forensic Research
Open Access

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Research Article

Forensic DNA Typing of Old Skeletal Remains Using AmpFlSTR® Identifiler® PCR Amplification Kit

Mian Sahib Zar1*, Ahmad Ali Shahid1, Muhammad Saqib Shahzad2, Kyoung-Jin Shin3, Hwan Young Lee3, Muhammad Israr1, Eun Hye Kim3, Zia Ur Rahman1, Tayyab Husnain1

1Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore, Pakistan.

2Department of Forensic Sciences, Institute of Molecular Biology and Biotechnology, University of Lahore, Pakistan.

3Department of Forensic Medicine, Yonsei University College of Medicine, Seoul South Korea.

*Corresponding Author:
Mian Sahib Zar
Centre of Excellence in Molecular Biology (CEMB)
University of the Punjab Lahore
87-West Canal Bank Road Thokar Niaz BaigLahore, Punjab, Pakistan
Tel: +92-333-9701436
E-mail: msahibzar@yahoo.com

Received date: December 06, 2013; Accepted date: December 24, 2013; Published date: December 28, 2013

Citation: Zar MS, shahid AA, Shahzad MS, Shin KJ, Lee HY, et al. (2013) Forensic DNA Typing of Old Skeletal Remains Using AmpFlSTR®Identifiler® PCR Amplification Kit. J Forensic Res 5:211. doi: 10.4172/2157-7145.1000211

Copyright: © 2013 Zar MS. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Background: In this study DNA typing of old skeletal remains was improved through AmpFlSTR®Identifiler®PCR amplification kit using different approaches. Methodology: DNA extraction was carried out by silica columns based total demineralization extraction method.DNA quantification was carried out by Real Time PCR. DNA amplification was carried out by using AmpFlSTR®Identifiler® PCR amplification kit with modified conditions. Capillary electrophoresis and Data analysis were carried out by Genetic Analyzer 3130 (ABI) and GeneMapper ID software version 3.2. Results: DNA was detected in 17 out of 24 skeletal remains. Among them, in 7 samples DNA was in the range of 1-10 pg/µL, in 4 samples it was in the range of 22-69 pg/µL and in 6 samples the DNA was in the range of >100 pg/ µL. The CT value (<30) of 40 cycles indicated that the PCR inhibitors were removed during DNA extraction method. Promising results were obtained by increasing the number of PCR cycles from standard 28 to 33 instead of 32 in PCR reaction. Finally it was observed that consensus approach produced reliable and reproducible DNA profiles from old skeletal remains. Conclusions: Forensic DNA typing of old skeletal remains, through a multiplex AmpFlSTR®Identifiler® PCR amplification kit, is improved by using: a highly effective DNA extraction method, modified and optimized PCR conditions, increasing sensitivity of PCR amplification and consensus approach

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