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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. W. Dwiputri Ade Ari Sundari Ade Ari Sundari Amelia Putri, Ni Luh Dian Saptari Arifani Siswidiasari Asmara A. A. R. Astuti, M.A.P. Ayu Nyoman Chandra Yustiana Ayu Putu Chika Iswari Anjani, Sang Cista Dewi, Ni Putu Fiona Dek Pueteri Dewi Suryani Deniariasih, N.W. Devi, Ni Wayan Antika Sri Devita Kusdianingrum Dewa Ayu Swastini Dewi Andayani Farmawati Dwicandra, N.M.O. Dyah Subadrika Warma Dewi Endang Kumolosasi Gede Bayu Surya Ekayana, I Heni Pujiastuti I Gede Ketut Sajinadiyasa I Gusti A. A. Santhi Rahmaryani I Gusti Ayu Agung Septiari I Ketut Juniarta I Ketut Tunas I Made Dira Swantara I Nengah Wirajana I Nyoman Gede Budiana I Wayan Kasa Ida Ayu Gendari Ida Ayu Ratih Dwi Nugraha Putri Ida Bagus Nyoman Putra Dwija Indra Juana Adikara Jennifer Tamara Jennifer Tamara Kadek Ratna Sari Dewi Kadek Widya Yuli Hartati Ketut Ratnayani Ketut Widyani Astuti Liangky Syane S Luh Vida Sasmitha Luk Ketut Budi Maitriani M. A. Pratiwi Made Dharmesti Wijaya Made Rai Dwitya Wiradiputra Marfu'ah, Nurul Marlia Singgih Wibowo Mirawati N.K.W. Ni Kadek Ari Cipta Pratiwi Ni Kadek Ariani Ni Ketut Sri Anggreni Ni Komang Sasi Ani Ni Made Febrianti Ni Made Yustikarini Ni Nyoman Pradnya Utari Ni Putu Ariantari Ni Putu Intan Satya Dewi Ni Putu Monica Rosdiana Dewi Paramitha Ni Wayan Sukma Pramitha Sari1 Osamu Iitsuka Pradnyaniti, D.G Putri, Ni Kadek Sri Adnyani Alit Putu Ayu Indrayathi Putu Lita Astriani Rasmaya Niruri Ratna Sari Dewi, Kadek Rini Noviyani Sang Ayu Putu Chika Iswari Anjani Saputra, Gusti Nyoman Oka Sari Dewi, Kadek Ratna Sudarma, I Wayan Ari Syamsul Arifin Tasya Pramiswari Tirtawati, Ni Putu Ayu Wijayakusuma, I Gusti Ngurah Lanang Yayanasri Yustiantara, Putu Sanna