OPTIMASI SUHU ANNEALING PROSES PCR AMPLIFIKASI GEN ATP1 A1 SAPI JABRES DAN GALEKAN

Optimization of Annealing Temperature PCR Process ATP1 A1 Gene Amplification in Jabres and Galekan Cows

Authors

  • Dewi Khosiya Robba Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Dyah Tuwi Ramsiati Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Wahyuni Indah Wulansari Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Mochammad Chanafi Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Asepriyadi Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Rina Ariyanti Pusat Riset Peternakan, Organisasi Riset Pertanian dan Pangan, Badan Riset dan Inovasi Nasional
  • Ullin Nihaya Departement of Information Management, Tatung University https://orcid.org/0000-0003-2733-891X

DOI:

https://doi.org/10.37090/jwputb.v8i1.1160

Abstract

Adanya kenaikan suhu pada lingkungan sekitar ternak sapi lokal Jabres dan sapi Galekan memiliki efek negatif terhadap perkembangan, pertumbuhan, produktivitas dan reproduksi-nya. Untuk itu, diperlukan metode untuk mengetahui kemampuan adaptasi sapi terhadap cekaman panas. Pada penelitian ini, sampel yang digunakan adalah DNA sapi lokal Jabres dan sapi Galekan dengan menggunakan metode penelitian molekuler in-vitro seperti isolasi DNA, Amplifikasi gen ATP1 A1 dengan Optimasi PCR, elektroforesis dan hasil visualisasinya. Isolasi DNA pada 8 sampel darah sapi Jabres dan sapi Galekan menunjukkan hasil isolasi DNA yang tepat dan sesuai dengan ditandai munculnya pita DNA yang terang dan tebal. Sedangkan, suhu annealing pada proses optimasi PCR yang digunakan untuk menempelkan primer forward ATP1 A1 5'- TCC CCA AGC TAG TGA CCA AG -3' dan primer reverse ATP1 A1 5'- TCT GTG GCT CTG ATT CTC CC -3' adalah 57oC, 57,3oC, 58,1oC, 59,3oC, 60,7oC, 61,9oC, 62,7oC, 63oC. Hasil optimasi PCR dielektroforesis dengan 100 volt selama 35 menit kemudian gel agarose hasil elektroforesis divisualisasikan menggunakan UV transilluminator sehingga didapatkan hasil suhu annealing yang optimal pada suhu 57 oC.

 

Kata Kunci : Sapi, PCR, gen ATP1A1, Annealing

References

Chen, J. M., Schütz, K. E., & Tucker, C. B. (2013). Dairy cows use and prefer feed bunks fitted with sprinklers. Journal of Dairy Science, 96(8), 5035–5045. https://doi.org/10.3168/jds.2012-6282

Chen, X., Shu, H., Sun, F., Yao, J., & Gu, X. (2023). Impact of Heat Stress on Blood, Production, and Physiological Indicators in Heat-Tolerant and Heat-Sensitive Dairy Cows. Animals, 13(16). https://doi.org/10.3390/ani13162562

Hidayati, H., Saleh, E., & Aulawi, T. (2016). Identifikasi keragaman gen bmpr-1b (bone morphogenetic protein receptor ib) pada ayam arab,ayam kampung dan ayam ras petelur menggunakan pcr-rflp. Jurnal Peternakan, 13(1), 1. https://doi.org/10.24014/jupet.v13i1.2383

Imran, S., Khan, M. S., Hassan, F. U., & Qureshi, Z. I. (2021). Genetic characterization of 38 cholistani breed of cattle for atp1a1 gene and its association to heat tolerance traits. Pakistan Journal of Agricultural Sciences, 58(1), 229–234. https://doi.org/10.21162/PAKJAS/21.11

Klabnik, J. L., Christenson, L. K., Gunewardena, S. S. A., Pohler, K. G., Rispoli, L. A., Payton, R. R., Moorey, S. E., Neal Schrick, F., & Edwards, J. L. (2022). Heat-induced increases in body temperature in lactating dairy cows: impact on the cumulus and granulosa cell transcriptome of the periovulatory follicle. Journal of Animal Science, 100(7). https://doi.org/10.1093/jas/skac121

Li, R., Ahmad, M. J., Hou, M., Wang, X., Liu, S., Li, J., Jiang, Q., Huang, J., & Yang, L. (2023). Identification of target genes and pathways related to heat tolerance in Chinese Holstein cows. Livestock Science, 271. https://doi.org/10.1016/j.livsci.2023.105213

Liberati, P. (2023). An Active Drying Sensor to Drive Dairy Cow Sprinkling Cooling Systems. Sustainability (Switzerland), 15(12). https://doi.org/10.3390/su15129384

Macavoray, A., Rashid, M. A., Rahman, H., & Shahid, M. Q. (2023a). On-Farm Water Use Efficiency: Impact of Sprinkler Cycle and Flow Rate to Cool Holstein Cows during Semi-Arid Summer. Sustainability (Switzerland), 15(4). https://doi.org/10.3390/su15043774

Macavoray, A., Rashid, M. A., Rahman, H., & Shahid, M. Q. (2023b). Sustainable Management of Dairy Cows in Semi-Arid Summers: Additional Early Nighttime Sprinkler Cooling for Heat Stress Mitigation. Sustainability (Switzerland), 15(15). https://doi.org/10.3390/su151511665

Marwayana, O. (2015). Ekstraksi Asam Deoksiribonukleat (DNA) dari Sampel Jaringan Otot. OSEANA, XI, 1–9.

Mavrogonatou, E., Papadimitriou, K., Urban, J., Papadopoulos, V., & Kletsas, D. (2015). Deficiency in the α1 Subunit of Na+/K+-ATPase Enhances the Anti-Proliferative Effect of High Osmolality in Nucleus Pulposus Intervertebral Disc Cells. Journal of Cellular Physiology, 230. https://doi.org/10.1002/jcp.25040

Osei-Amponsah, R., Dunshea, F. R., Leury, B. J., Cheng, L., Cullen, B., Joy, A., Abhijith, A., Zhang, M. H., & Chauhan, S. S. (2020). Heat stress impacts on lactating cows grazing australian summer pastures on an automatic robotic dairy. Animals, 10(5). https://doi.org/10.3390/ani10050869

Prasanna, J. S., Rao, S. T. V., Prakash, M. G., Rathod, S., Kalyani, P., & Reddy, B. R. (2022). Association of SSCP Polymorphisms of HSP70 Gene with Physiological, Production and Reproduction Performance in Sahiwal and Crossbred Cows. Asian Journal of Dairy and Food Research, 41(2), 150–155. https://doi.org/10.18805/ajdfr.DR-1796

Prakoso, S. P., Wirajana, I. N., & Suarsa, I. W. (2016). Amplifikasi fragmen gen 18s rRNA pada dna metagenomik madu dengan teknik PCR (polymerase chain reaction). Indonesian Journal of Legal and Forensic Sciences (IJLFS), 7(3), 1. https://doi.org/10.24843/ijlfs.2017.v07.i01.p03

Rajamanickam, G. D., Kastelic, J. P., & Thundathil, J. C. (2017). Na/K-ATPase regulates bovine sperm capacitation through raft- and non-raft-mediated signaling mechanisms. Molecular Reproduction and Development, 84(11), 1168–1182. https://doi.org/10.1002/mrd.22879

Robba, D.K., Ramsiati,D.T., Wulansari,W.i. Isolasi DNA Darah Sapi Madura dengan Modifikasi Metode KIT. Seminar Nasional IV Hasil-hasil Penelitian dan Pengabdian kepada Masyarakat. Fakultas Peternakan Universitas Jambi. 7 November 2022. 4(1), 58-63.

Romjali, E. (2018). Program pembibitan sapi potong lokal Indonesia. Wartazoa, 28(4), 199–210.

Shpigel, N. Y., Pasternak, Z., Factor, G., & Gottlieb, Y. (2015). Diversity of bacterial biofilm communities on sprinklers from dairy farm cooling systems in Israel. PLoS ONE, 10(9). https://doi.org/10.1371/journal.pone.0139111

Sutedjo, H. (2016). Dampak fisiologis dari cekaman panas pada ternak (physiology effect of heat stress on animal). 3(1), 93–105.

Tresoldi, G., Schütz, K. E., & Tucker, C. B. (2018). Cooling cows with sprinklers: Timing strategy affects physiological responses to heat load. Journal of Dairy Science, 101(12), 11237–11246. https://doi.org/10.3168/jds.2018-14917

Yuwono. 2005. Teori dan Aplikasi PCR. Yogyakarta: Erlangga.

Zamorano-Algandar, R., Medrano, J. F., Thomas, M. G., Enns, R. M., Speidel, S. E., Sánchez-Castro, M. A., Luna-Nevárez, G., Leyva-Corona, J. C., & Luna-Nevárez, P. (2023). Genetic Markers Associated with Milk Production and Thermotolerance in Holstein Dairy Cows Managed in a Heat-Stressed Environment. Biology, 12(5). https://doi.org/10.3390/biology12050679

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01-03-2024

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Dewi Khosiya Robba, D. Tuwi Ramsiati, W. Indah Wulansari, M. . Chanafi, Asepriyadi, R. Ariyanti, and U. Nihaya. “OPTIMASI SUHU ANNEALING PROSES PCR AMPLIFIKASI GEN ATP1 A1 SAPI JABRES DAN GALEKAN: Optimization of Annealing Temperature PCR Process ATP1 A1 Gene Amplification in Jabres and Galekan Cows”. Wahana Peternakan, vol. 8, no. 1, Mar. 2024, pp. 10-19, doi:10.37090/jwputb.v8i1.1160.

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Vol. 8 No. 1 (2024): Wahana Peternakan

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Copyright (c) 2024 Dewi Khosiya Robba, Dyah Tuwi Ramsiati, Wahyuni Indah Wulansari, Mochammad Chanafi, Asepriyadi, Rina Ariyanti, Ullin Nihaya

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