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Abstract
The aim of this research was to detect the skin which comes from goat and pig using PCR-RFLP method so the raw material used in rambak product was discovered. The comparison of combination between goat and pig skin was 100, 75:25, 91:9, 94:6, 97:3, and 99:1. PCR-RFLP uses the universal primary from from mitochondria cytochrome b gen which results in the length of fragment of 359 bp. The restriction enzyme used in the DNA cut is BamHI enzyme and BseDI enzyme. The result of the research showed that seven samples have been successfully isolated perfectly so the total bands of genomic DNA have been obtained which is clearly seen and amplification with target of cytochrome b gen results in PCR product of goat and pig of 359 bp. The digestion's result using BamHI enzyme gets fragment size on 359 bp in length on goat samples and length of fragment size of 359, 244, and 115 bp on samples which contain pig. The digestion results of using BseDI results in fragment size of 359 bp in length on goat samples and the fragment size of 359, 228, and 131 bp in length on samples which contain pig. The conclusion of the research is PCR-RFLP using BamHI enzyme and BseDI enzyme can be used to detect the types of pig skin, but can't be used to detect the goat skin.
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References
- Erwanto, Y., M.Z. Abidin, A. Rohman, and Sismindari. 2011. PCR-RFLP using BseDI enzyme for pork authentication in sausage and nugget products. Media Peternakan. 13(1): 14-18.
- Erwanto, Y., M.Z. Abidin, Sismindari, and A. Rohman. 2012. Pig species identification in meatballs using polymerase chain reaction-restriction fragment length polymorphism for halal authentication. International Food Research Journal 19:901-906.
- Fauziah, A.R. 2016. Identification of skin contaminants pork on rambak crackers with method real-time polymerase chain reaction using d-loop 22 'mitochondrial primer for Master Thesis, Universitas Gadjah Mada, Yogyakarta.
- Fitrianingsih. 2013. Optimization of DNA isolation on processed meat as the basis for detection of porcine contamination for master thesis, University of Gajah Mada, Yogyakarta.
- Grifiiths, A.J.F., J.H. Miller, D.T. Suzuki, R.C. Lewontin and W.M Gelbert. 1998. An Introduction to Genetic Analysis. Six Edition. W.H. Freeman and Company. New York.
- Jonasi, A. 2006. Species specific detection of meat by polymerase chain reaction techniques for master thesis, University Corvinus, Budapest.
- Kocher, Z., A. S.V. Thomas, S. Meyer, Edwards, and F.X. Paabo. 1989. Dynamic of mitochondrial DNA evolution in animal: amplifications and sequencing with conversed primers. Proceedings of the National Academy of Science of the USA. 86: 6169-6200.
- Murugaiah, C., Z.M. Noor, M. Mastakim, L.M., Bilung, J. Selamat and S. Radu. 2009. Meat species identification and halal authentication analysis using mitochondrial DNA. Meat Sci. 83: 57-61.
- Ong S.B., M.I. Zuraini, M. G. Jurin, Y.H. Cheah, R. Tunung, L.C. Chai, Y. Haryani, F.M. Ghazali and R. Son. 2007. Meat moleculer detection sensitivity of PCR-RFLP in species differentiation of meat from animal origin. ASEAN Food Journal 14: 51-59.
- Sambrook, J., E.F. Fritch and T. Maniatis. 1989. Molecular Cloning, A. Laboratory Manual. 2nded. Cold SpringHarbour Lab. Press, New York.
- Sarkar, K.T. 1982. Processing of goat skins for commercial leathers. Published by K.T. Sarkar. Madras, India. pp. 1-5.
- Trijoko, R., W. Cahyaningtyas, Surajiman, Istini, and D. Pranowo. 2012. Validation of PCR-RFLP testing method to detect porcine contamination in chicken nugget. Indo. J. Chem. 12: 302-307.
- Watson, D., M. James, J. Gilman, Witkowski and M. Zoller. 1992. Recombinan DNA. 2nded. United States of America.
References
Erwanto, Y., M.Z. Abidin, A. Rohman, and Sismindari. 2011. PCR-RFLP using BseDI enzyme for pork authentication in sausage and nugget products. Media Peternakan. 13(1): 14-18.
Erwanto, Y., M.Z. Abidin, Sismindari, and A. Rohman. 2012. Pig species identification in meatballs using polymerase chain reaction-restriction fragment length polymorphism for halal authentication. International Food Research Journal 19:901-906.
Fauziah, A.R. 2016. Identification of skin contaminants pork on rambak crackers with method real-time polymerase chain reaction using d-loop 22 'mitochondrial primer for Master Thesis, Universitas Gadjah Mada, Yogyakarta.
Fitrianingsih. 2013. Optimization of DNA isolation on processed meat as the basis for detection of porcine contamination for master thesis, University of Gajah Mada, Yogyakarta.
Grifiiths, A.J.F., J.H. Miller, D.T. Suzuki, R.C. Lewontin and W.M Gelbert. 1998. An Introduction to Genetic Analysis. Six Edition. W.H. Freeman and Company. New York.
Jonasi, A. 2006. Species specific detection of meat by polymerase chain reaction techniques for master thesis, University Corvinus, Budapest.
Kocher, Z., A. S.V. Thomas, S. Meyer, Edwards, and F.X. Paabo. 1989. Dynamic of mitochondrial DNA evolution in animal: amplifications and sequencing with conversed primers. Proceedings of the National Academy of Science of the USA. 86: 6169-6200.
Murugaiah, C., Z.M. Noor, M. Mastakim, L.M., Bilung, J. Selamat and S. Radu. 2009. Meat species identification and halal authentication analysis using mitochondrial DNA. Meat Sci. 83: 57-61.
Ong S.B., M.I. Zuraini, M. G. Jurin, Y.H. Cheah, R. Tunung, L.C. Chai, Y. Haryani, F.M. Ghazali and R. Son. 2007. Meat moleculer detection sensitivity of PCR-RFLP in species differentiation of meat from animal origin. ASEAN Food Journal 14: 51-59.
Sambrook, J., E.F. Fritch and T. Maniatis. 1989. Molecular Cloning, A. Laboratory Manual. 2nded. Cold SpringHarbour Lab. Press, New York.
Sarkar, K.T. 1982. Processing of goat skins for commercial leathers. Published by K.T. Sarkar. Madras, India. pp. 1-5.
Trijoko, R., W. Cahyaningtyas, Surajiman, Istini, and D. Pranowo. 2012. Validation of PCR-RFLP testing method to detect porcine contamination in chicken nugget. Indo. J. Chem. 12: 302-307.
Watson, D., M. James, J. Gilman, Witkowski and M. Zoller. 1992. Recombinan DNA. 2nded. United States of America.