Determination of Cattle and Buffalo Skin Crackers Using Polymerase Chain Reaction Restriction Fragment Length Polymorphism

Rulli Riana Dewi(1*), Yuny Erwanto(2), Nanung Agus Fitriyanto(3)

(1) Fakultas Peternakan Universitas Gadjah Mada
(2) Fakultas Peternakan Universitas Gadjah Mada
(3) Fakultas Kedokteran Hewan Universitas Gadjah Mada
(*) Corresponding Author


The aim of this study was to determine of cattle and buffalo species based on cytochrome b gene using PCR-RFLP. Cattle and buffalo hides were obtained from a slaughterhouse in Yogyakarta and Kudus Regency. To confirm the effectiveness and specificity of this fragment, there are seven of DNA mixture samples in various levels. Isolate DNA samples were amplified using universal primer of cytochrome b gene, then PCR amplicon was digested by RsaI restriction enzyme.. The result showed that mitochondrial cytochrome b gene successfully amplified fragments of 359 bp. RsaI restriction enzyme was able to cleave buffalo cytochrome b gene into two fragment  (326 and 23 bp), while the cytochrome b gene of the skin cattle DNA was uncleaved. . In conclusion, this study indicated that mixture DNA of cattle and buffalo hides could be digested by RsaI restriction enzyme  and determination of the buffalo hides in mixture samples could be detected into  10% level. Furthermore, RsaI enzyme could be used to specific identification buffalo species. PCR-RFLP technology has a potential and reliable method to identify  of the existence of r buffalo hides in the mixture with other hides.


Cattle and buffalo hides; Cytochrome b; Crackers; Polymerase chain reaction

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Bromley, C., and M. Maher-Sturm. (2005). The polymerase chain reaction (PCR): solution PCR on paraffin-embedded human tissues. The J. Histotechnol. 28 (4): 219-222.

Caine, L., G. Lima, L. Pontes, D. Abrantes, M. Pereira, and M. F. Pinheiro. (2006). Species identification by cytochrome b gene: Casework samples. Elsevier. 145-147.

Couto, M. C. M., A. P Sudre, M. F. Lima, and T. C. B. Bomfim. (2013). Comparison of techniques for DNA extraction and agarose gel staining of DNA fragments using samples of Cryptosporidium. J. Vet. Med. 58 (10): 535-542.

Doosti, A.; Ghasemi Dehkordi, P.; Rahimi, E. (2011). Molecular assay to fraud identification of meat products. Journal of Food Science and Technology, doi:10.1007/s13197-011-0456-3Erwanto, Y., M. Z. Abidin, A. Rohman, and Sismindari. 2011. PCR-RFLP using BseDI enzyme for pork authentication in sausage and nugget product. J. Anim. Sci. and Tech. 34 (1): 14-18.

Erwanto, Y., M. Z. Abidin, E. Y. P. Muslim, Sugiyono, and A. Rohman. (2014). Identification of pork contamination in meatballs of Indonesia local market using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Asian Autralas. J. Anim. Sci. 27(10): 1487-1492.

Hartatik, T., S. D. Volkandari, Sumadi, and Widodo. (2013). The application of polymerase chain reaction – restriction fragment polymorphisms (PCR-RFLP) to determine genetic diversity of Madura cattle in Sapudi island. I. J. Biotech. 18 (1): 70-74.

Kocher, T. D., A. Thomas, S. V. Meyer, S. Edwards, & F. X. Paabo. 1989. Dymanic of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proc. Nat. Acad. of Sci.. 86: 6169-6200.

Kumar, R. A., D. J. Odenburg, and A. J. Bendich. (2014). Changes in DNA damage, molecular integrity, and copy number for plastid DNA and mitochondrial DNA during maize development. J. Exp. Bot. 65 (22): 6425-6439.

Muttaqien, A. T., Y. Erwanto, and A. Rohman. (2016). Determination of buffalo and pig “Rambak” crackers using FTIR spectroscopy and chemometrics. Asian J. Anim. Sci. 1-10.

Sambrook, J., D. W. Russel, and T. Maniatis. (2001). Molecular Cloning: A Laboratory Manual 3rd Edition. Cold Spring Harbor Laboratory Press. New York.

Srisakwattana, K., C. Pansin, S. Chethasing, K. Tasripoo, W. Nualchuen, S. Usawang, and M. Kamonpatana. (2016). PCR-RFLP of mitochondrial DNA of swamp buffaloes in breeding station. Ital. J. Anim. Sci. 6 (2): 334-337.

Wu, Z., I. Nagano, E. Pozio, and Y. Takahashi. (1999). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for identification of Trichinella isolates. Parasitology. 118: 211-218.

Xu, K., J. F. X. Ma, X. Wang, D. Zhou and Z. Dai. (2015). Identification of tuna species (Thunnini tribe) by PCR-RFLP analysis of mitochondrial DNA fragments. Food and Agric. 27 (3): 301-313.


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