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Journal : Jurnal Veteriner

Optimasi Suhu Annealing Tiga Regio Berbeda Isolat Multidrug Resistance Mycobacterium Tuberculosis dengan Metode Multiplex Polymerase Chain Reaction (ANNEALING TEMPERATURE OPTIMIZATION ON THREE DIFFERENT REGIONS OF MYCOBACTERIUM TUBERCULOSIS MULTIDRUG RESI Indra Juana Adikara; I Nengah Wirajana; Sagung Chandra Yowani
Jurnal Veteriner Vol 17 No 4 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Multiplex PCR is a method used to amplify more than one target sequences simultaneously. The aimof this research was to optimize PCR on the region of inhA promoter, inhA gene and katG gene usingMultidrug Resistance Tuberculosis (MDR-TB) Isolate. Isolation of DNA was done by using High Pure PCRTemplate Preparation Kit. Amplification process was done by Multiplex PCR usingthree pairs primers i.e.mabA-inhA-promoter-FS and mabA-inhA-promoter-R,inhA (F) and inhA (R) and KG24F and KG60R.Amplification process started by predenaturation at 95°C for 15 minutes, followed by 45 cycles consistingof denaturation at 94°C for 1 minute, annealing at 56°C, 57°C and 58°C for 1 minute and 20 seconds andextension at 72°C for 2 minutes. Then it is finished by postextension at 72°C for 10 minutes. PCR productwas detected by electrophoresis and visualized under UV Transiluminator. Annealing temperature of56°C resulted in a thicker, clearerandaccording to the desired size as compared to that of with 57°C and58°C. Conclusion that obtained was annealing temperature 56°C was optimum annealing temperatureon inhA promoter, inhA gene and katG gene region of Mycobacterium tuberculosis Multidrug Resistanceisolate using Multiplex Polymerase Chain Reaction.
Co-Authors A. A. B. Putra Anak Agung Istri Agung Mayun Laksmiwati D. Rizkiyanti Darma Asih Yuliana Dewi Andayani Farmawati Dewi, I Gusti Ayu Agung Gangga Samala Diah Suci Eddy Bagus Wasito Gusti A Malelak H.M. Soekry Erfan Kusuma I Gede Mahardika I Gusti Ngurah Bagus Andre Hartawan I Ketut Gede Dharma Dewantara I Made Agus Gelgel Wirasuta I P. J. D. A. Suartama I Putu Mahendra I W BUDIARSA SUYASA I Wayan Gede Gunawan I Wayan Suarsa I. A. Gede Widihati Ida Ayu Ratih Dwi Nugraha Putri Ida Bagus Amertha Putra Manuaba Ida Bagus Putra Manuaba Ida Bagus Putu Eristya Putra Ika Kurniawati Indra Juana Adikara Inten Hardianti Nizar Iryanti Eka Suprihatin James Sibarani Kazuo Sakka Ketut Ratnayani Khisan Qamariya Kimura, Tetsuya Kimura, Tetsuya Kusuma, Soekry Erfan Kusuma, Soekry Erfan Luh De Dwi Jayanthi Luk Ketut Budi Maitriani M. Manurung Made Arsa Made Dharmesti Wijaya Made Rai Dwitya Wiradiputra Mirawati N.K.W. N. M. T. Juliasari N. W. Bogoriani Ni Komang Ariati Ni Komang Lia Wahyuni Ni Luh Md. Widayantini Ni Luh Putu Mustika Praptiwi Ni Made Suaniti Ni Made Yustikarini Ni Nengah Kartini Asih Ni Nyoman Tri Puspaningsih Ni Putu Ariantari Ni Putu Citra Anggryni Sugitha Ni Putu Frida Oktaningtias Widiarthi Ni Putu Rahayu Artini O. Ratnayani Oka Ratnayani P. Suarya Pradnyaniti, D.G Putra, Pramana Kumala Putri, Ni Wayan Prasanthi Swarna Putu Elvira Yulianthi Putu Suarya R. R. Sirait Ro’yal Aini Sagun Chandra Yowani Sakka, Kazuo Sakka, Kazuo Satriya Putra Prakoso Soekry Erfan Kusuma Tetsuya Kimura Wahyu Dwijani Sulihingtyas Y. H. Pratiwi