Permana, Jonathan Arsha (2022) Isolasi Dan Identifikasi Bakteri Baru Dari Lahar Semeru Menggunakan Metode Pewarnaan Gram Dan Analisis Gen 16S rRNA. Other thesis, Institut Teknologi Sepuluh Nopember.
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Abstract
Eksplorasi terhadap mikroorganisme ekstremofilik di Kawasan Gunung Semeru masih sangat minim dilakukan. Penelitian ini bertujuan untuk melakukan isolasi terhadap isolat bakteri yang terkandung di dalamnya dan melakukan identifikasi menggunakan pewarnaan gram serta analisis gen 16S rRNA. Hasil isolasi diperoleh bakteri dengan diberi kode LS-ISP2-JS2 lalu diidentifikasi menggunakan metode pewarnaan gram serta analisis gen 16S rRNA. Hasil dari pewarnaan gram menunjukkan bahwa isolat merupakan gram negatif dan memiliki bentuk basil. DNA isolat diekstraksi dan diampifikasi gen 16S rRNA dengan forward primer 27F (5’-AGAGTTTGATCMTGGCTCAG-3’) dan reverse primer 1492R (5’-TACGGYTACCTTGTTACGACTT-3’) menggunakan metode PCR. Proses elektroforesis menggunakan gel agarose 2% menunjukkan pita tunggal dengan ukuran ± 200 bp. Data dari sekuensing menunjukkan jumlah urutan basa nukleotida sebanyak 146 basa. Sekuen nukleotida hasil sekuensing dibandingkan dengan data pada National Center for Biotechnology Information (NCBI) dengan menggunakan metode BLAST. Pembuatan pohon filogenetik yang dilakukan menggunakan software MEGA11 menunjukkan hasil bahwa isolat LS-ISP2-JS2 memiliki kemiripan dengan Uncultured bacterium clone 1 16S ribosomal RNA gene dengan presentase kemiripan sebesar 98,56%, yang mengindikasikan bahwa isolat tersebut merupakan bakteri yang belum pernah dikulturkan di luar lingkungannya dan dapat dikategorikan sebagai bakteri baru.
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Exploration of extremophilic microorganisms in Mount Semeru area is rarely done. This research was conducted to isolate the bacterial isolates contained in it and to identify using gram staining and analysis of the 16S rRNA gene. The results of the isolation obtained were bacteria coded LS-ISP2-JS2 and then identified using the gram staining method and analysis of the 16S rRNA gene. The result of gram staining showed that the isolate was gram negative and had a bacillus form. The isolate DNA was extracted and amplified 16S rRNA gene with forward primer 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and reverse primer 1492R (5'-TACGGYTACCTTGTTACGACTT-3') using PCR method. The electrophoresis process that using 2% agarose gel showed a single band with a size of ± 200 bp. The data from the sequencing shows the number of nucleotide base sequences is 146 bases. The sequenced nucleotide were compared with the data on NCBI (National Center for Biotechnology Information) using the BLAST method. Phylogenetic tree is made using MEGA11 software, showed that the LS-ISP2-JS2 isolate had similarities with Uncultured bacterium clone 1 16S ribosomal RNA gene with a similarity percentage of 98.56%, which indicated that the isolate was an uncultured bacterium and can be categorized as a novel bacteria.
| Item Type: | Thesis (Other) |
|---|---|
| Additional Information: | RSKi 589.9 Per i-1 2022 |
| Uncontrolled Keywords: | Semeru, Ekstermofilik, Gen 16S rRNA, Pewarnaan Gram, Bakteri Baru. Semeru, Extremophilic, 16S rRNA Gene, Gram Staining, Novel Bacteria. |
| Subjects: | Q Science > QR Microbiology > QR74.8 Bacteria |
| Divisions: | Faculty of Science and Data Analytics (SCIENTICS) > Chemistry > 47201-(S1) Undergraduate Thesis |
| Depositing User: | Mr. Marsudiyana - |
| Date Deposited: | 06 May 2026 04:12 |
| Last Modified: | 06 May 2026 04:12 |
| URI: | http://repository.its.ac.id/id/eprint/133008 |
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