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All Journal METAMORFOSA Journal of Biological Sciences
Ida Ayu Ratih Dwi Nugraha Putri
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology

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OPTIMASI DIGESTI ENZIM RESTRIKSI SacII PADA ISOLAT Mycobacterium tuberculosis H37Rv UNTUK DETEKSI MUTASI PROMOTER inhA PADA KASUS MDR-TB DENGAN METODE PCR-RFLP Ida Ayu Ratih Dwi Nugraha Putri; Sagung Chandra Yowani; I Nengah Wirajana
Metamorfosa: Journal of Biological Sciences Vol 4 No 1 (2017)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2017.v04.i01.p14

Abstract

The aim of this study was to optimize the digestion process of SacII restriction enzyme in Mycobacterium tuberculosis H37Rv isolate. SacII enzyme that have CCGC^GG restriction site could be used to detect the mutation at -24 promoter region of inhA M. tuberculosis. The mutation in this position is one of the causes of isoniazid resistance. The methods of this study were conducted in several step, respectively: DNA isolation of M. tuberculosis H37Rv, amplification of inhA promoter region of M. tuberculosis H37Rv using PCR method, and digest optimization using SacII. In this study, optimization of SacII digestion was conducted for formulaion and incubation time. The formulation was optimized by GM-ONE Buffer and GM-Buffer IV, meanwhile the incubation time was optimized in 1 hour, 2 hour, and 3 hour. The result of this study showed that SacII work optimally in GM-Buffer IV compared to GM-ONE Buffer and incubate for 3 hours. Under the optimal digest condition, SacII enzyme could be used to detect the mutation at -24 promoter region of inhA in the case of MDR-TB using PCR-RFLP method.
Co-Authors I Nengah Wirajana Sagun Chandra Yowani