Comparative Study of Sperm DNA Isolation Method for Forensic Analysis

  • Patricia Gita Naully Department of Medical Laboratory Technology (D-4), Stikes Jenderal Achmad Yani Cimahi. West Java, Indonesia


Forensic is a multi-discipline science that is used to obtain evidence of various criminal cases, such as rape. DNA analysis on sperm specimen is needed to identify the rapist. However, the success of this analysis depends on the DNA isolation method used. Several methods of DNA isolation from human sperm have been developed, but no method has been proven effective for the forensic analysis need. This study aimed to determine the effective sperm DNA isolation method for forensic analysis. In this study, the DNA of sperm specimens was isolated using three methods: Boiling Water, modified TRIzol, and Chelex-100. The DNA isolation result was visualized using agarose gel electrophoresis method. The concentration and purity of isolated DNA were measured using a Nanodrop by comparing the absorbance of DNA at λ 260 nm and protein at λ 280 nm. The effectiveness of the sperm DNA isolation method was determined based on the concentration and purity of DNA, the specimen volume, the implementation time, and the costs involved. The result showed that the successful methods for isolating sperm DNA were TRIzol and Chelex-100. The quantity of DNA isolated using the modified TRIzol method was 1,5 times higher than Chelex-100 but equired 120 times more specimen volume than Chelex-100. From 25 µl sperm specimens, the concentration of DNA isolated using the Chelex-100 method was 612.6 ng/µl with a purity of about 1.7. Therefore, it can be concluded that the Chelex-100 is the most effective method for isolating sperm DNA for forensic analysis.

Keywords: boiling water, chelex-100, DNA isolation, sperm, trizol


Alarcón-Zúñiga, B., Zepeda-Batista, J.L., Ruíz-Flores, A., Gómez-Meza, L.J., García-Muñiz, J.G., Núñez-Domínguez, R., Ramírez-Valverde, R., Villegas-Velázquez, I., 2016. Modificación del método de tiocianato de guanidina para extraer ADN de semen para análisis genómico en mamíferos. RMCP 7, 405.
Anvar, Z., Namavar-Jahromi, B., Ebrahimi, S., Gharesi-Fard, B., 2015. Genomic DNA Extraction from Sperm. Journal of Advanced Medical Sciences and Applied Technologies 1, 120.
Biswas, K., Garg, N., Biswas, R., 2017. PCR-RFLP of Calotropis gigantea (L)- A Tool for Forensic Application 12, 61–67.
Darbandi, M., Darbandi, S., Khorram Khorshid, H.R., Akhondi, M.M., Mokarram, P., Sadeghi, M.R., 2018. A simple, rapid and economic manual method for human sperm DNA extraction in genetic and epigenetic studies. Middle East Fertility Society Journal 23, 216–219.
Dashti, A.A., Jadaon, M.M., Abdulsamad, A.M., Dashti, H.M., 2009. Heat Treatment of Bacteria: A Simple Method of DNA Extraction for Molecular Techniques. KUWAIT MEDICAL JOURNAL 7.
Jehuda, V., 2013. Ekstraksi DNA dari Sperma pada Kondom dan Kain yang Tersimpan Sampai Dua Belas Hari 1, 28–39.
Lorenz, T.C., 2012. Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies. J Vis Exp.
Manuja, A., 2010. Evaluation of Different Methods of DNA Extraction from Semen of Buffalo (Bubalus Bubalis) Bulls. Buffalo Bulletin 8.
M.S. Lanang, A.A.G., Junitha, I.K., Suaskara, I.B.M., 2013. Ekstrasksi DNA Sperma pada Kondom yang Disimpan dalam Rentang Waktu Berbeda. Jurnal Biologi 17, 47–50.
Narejo, N.B., Avais, M.A., 2012. SINDH UNIVERSITY RESEARCH JOURNAL (SCIENCE SERIES). Sindh Univ. Res. Jour. 44, 251–254.
Queipo-Ortuño, M.I., De Dios Colmenero, J., Macias, M., Bravo, M.J., Morata, P., 2008. Preparation of Bacterial DNA Template by Boiling and Effect of Immunoglobulin G as an Inhibitor in Real-Time PCR for Serum Samples from Patients with Brucellosis. CVI 15, 293–296.
Rio, D.C., Ares, M., Hannon, G.J., Nilsen, T.W., 2010. Purification of RNA Using TRIzol (TRI Reagent). Cold Spring Harb Protoc 2010, pdb.prot5439.
Roux, C., Crispino, F., Ribaux, O., 2012. From Forensics to Forensic Science. Current Issues in Criminal Justice 24, 7–24.
Sandwinata, Muh.F., 2019. Analisis DNA Dalam Kasus Forensik. TEKNOSAINS 12.
Schiebelhut, L.M., Abboud, S.S., Daglio, L.E.G., Swift, H.F., Dawson, M.N., 2017. A comparison of DNA extraction methods for high-throughput DNA analyses. Molecular Ecology Resources 17, 721–729.
Silva, G.A. da, Bernardi, T.L., Schaker, P.D.C., Menegotto, M., Valente, P., 2012. Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification. Braz. arch. biol. technol. 55, 319–327.
Ugale, B., Singh, D., Nagaraja, C.M., 2015. Temperature dependent structural variation from 2D supramolecular network to 3D interpenetrated metal–organic framework: In situ cleavage of S–S and C–S bonds. Journal of Solid State Chemistry 226, 273–278.
Walsh, P.S., Metzger, D.A., Higuchi, R., 2013. Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material. BioTechniques 54.
Wu, H., de Gannes, M.K., Luchetti, G., Pilsner, J.R., 2015. Rapid method for the isolation of mammalian sperm DNA. BioTechniques 58.
How to Cite
Naully, P. (2020). Comparative Study of Sperm DNA Isolation Method for Forensic Analysis. Biomedika, 13(2), 111-116.