ISOLASI DNA METAGENOMIK DARI SPUTUM PASIEN TUBERKULOSIS DAN AMPLIFIKASI DENGAN PRIMER PROMOTER inhA
Abstract
ABSTRAK: Penelitian ini bertujuan untuk memperoleh DNA metagenomik dari sputum pasien tuberkulosis dan mengamplifikasi dengan menggunakan primer promoter inhA. Penelitian ini dilakukan dalam tiga tahap, yaitu: isolasi DNA metagenomik dari sputum pasien tuberkulosis, amplifikasi menggunakan sepasang primer promoter inhA dari M. tuberculosis dengan metode PCR, dan elektroforesis gel agarosa terhadap hasil amplifikasi. Elektroforegram hasil amplifikasi menunjukkan bahwa isolasi DNA metagenomik dari sputum pasien tuberkulosis telah berhasil dilakukan dengan metode modifikasi maupun dengan kit. Ukuran pita fragmen DNA sekitar 284 bp dari hasil amplifikasi (amplikon) yang diperoleh dari DNA metagenomik sputum P.48B, P.46B, dan P.47B MDR- TB merupakan ukuran yang sesuai dengan bagaian dari daerah promoter inhA M. tuberculosis. Ukuran amplikon ini sama dengan ukuran amplikon yang sebelumnya telah diperoleh dari amplifikasi terhadap DNA M. tuberculosis oleh peneliti sebelumnya dengan menggunakan primer yang sama.
ABSTRACT: The aims of this research were to obtain metagenomic DNA from sputum of tuberculosis patients and to amplify it by using inhA promoter primer. This research was conducted in three steps covering the DNA metagenomic isolation from sputum of tuberculosis patients, amplification of inhA promoter region of M. tuberculosis by using specific primer pair by PCR (Polymerase Chain Reaction) method, and electrophoresis of amplified products using agarose gel. The electrophoregram of amplified products showed that the metagenomic DNA isolation from sputum of tuberculosis patients was successfully carried out both by the modified method and by a kit. The size of DNA fragment bands about 284 bp of amplified products (amplicons) which obtained from the metagenomic DNAs of P.48B, P.46B, P.47B sputum was suitable size with a part of inhA promoter region of M. tuberculosis. The size of these amplicons was the same size with the amplicons from M. tuberculosis DNA reported by other researchers.
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References
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