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All Journal METAMORFOSA Journal of Biological Sciences INDONESIAN JOURNAL OF MEDICAL LABORATORY SCIENCE AND TECHNOLOGY Bioscientist : Jurnal Ilmiah Biologi
Muhammad Taufiq Hidayat
Department of Medical Laboratory Technology, FKes, Nahdlatul Ulama University of Surabaya
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Health Professions Immunology & microbiology Social Sciences

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Modified Spin Column-Based RNA Extraction Methods of Staphylococcus aureus using PureLink® RNA Mini Kit and Basic Laboratory Instrument Muhammad Taufiq Hidayat; Endry Nugroho Prasetyo
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 3 No 2 (2021): Molecule analysis to advance laboratory diagnosis
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v3i2.1863

Abstract

RNA extraction is an important process before gene expression assessment at the transcriptomic level. RNA is a sensitive material to environmental factors such as temperature and contaminants, so the RNA extraction process generally requires sophisticated and expensive laboratory instruments. In this study, we extract RNA from Staphylococcus aureus bacteria using the PureLink® RNA Mini Kit. The instruments used in this study are basic instruments such as a hand homogenizer and non-thermal centrifuge. The results of RNA extraction were visualized using agarose gel electrophoresis. These results indicate that bacterial RNA extraction can be performed using the PureLink® RNA Mini Kit even with inexpensive basic laboratory instruments.
Comparison of Three DNA Isolation Methods of Aspergillus Niger Adyan Donastin; Maharani Pertiwi Koentjoro; Muhamad Taufik Hidayat; Endry Nugroho Prasetyo
Metamorfosa: Journal of Biological Sciences Vol 9 No 1 (2022)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2022.v09.i01.p07

Abstract

Polymerase Chain Reaction (PCR) is a technique that is applied to detect and test the presence of genetic material from pathogenic fungi. This method was further developed because it has a high sensitivity compared to the culture method in detecting the presence of pathogenic fungi. To perform sensitive, specific and reliable PCR-based assays, the availability of pure DNA as well as an easy-to-perform DNA extraction protocol is essential. Currently, existing protocols for DNA extraction generally require specialized kits and with the addition of enzymes. Therefore, this study was conducted to compare the quantity and quality of Aspergillus niger DNA isolation with three different methods. The stages of the research carried out included pure A. niger culture and total DNA isolation using three methods, namely the protocol according to the Wizard® Genomic DNA Purification Kit (P1) instructions, Modified Wizard® Genomic DNA Purification Kit (P2) and Monarch Genomic DNA Purification Kit NEB (P3). The results of the evaluation of DNA isolation using a spectrophotometer at a wavelength of A260/280 nm showed a ratio of 1.4, 1.8, and 1.7 at P1 and P2, and P3 respectively. The quantity obtained ranged from 210 to 305 ng/µL. The total DNA results obtained were then used for PCR testing of ITS ribosomal DNA fragments using ITS1 and ITS4 primers. Electrophorosis observation of PCR results showed that all samples produced bands with a length of 500 bp. The results of DNA isolation in the P1 method did not show bands on the agarose gel, but could be seen in the PCR product. Methods P2 and P3 showed DNA that had good quality and quality. The P2 and P3 methods use a cell destruction technique with liquid nitrogen and a combination of addition of proteinase K. The obtained protocol is expected to provide fast and good method information in the isolation of total DNA from A. niger. Keyword: Aspergillus niger, total DNA isolation, ITS1, ITS4
Antimicrobial Activity Test of Medicinal Plant Extract Using Antimicrobial Disc and Filter Paper Muhammad Taufiq Hidayat; Maharani Pertiwi Koentjoro; Endry Nugroho Prasetyo
Bioscientist : Jurnal Ilmiah Biologi Vol 11, No 1 (2023): June
Publisher : Department of Biology Education, FSTT, Mandalika University of Education, Indonesia.

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33394/bioscientist.v11i1.7544

Abstract

Plant extracts contain secondary metabolites such as flavonoids and phenolic compounds with antibiotic activity. Antibiotic activity test can be done by using the Kirby-Bauer disc method which the antibiotic activity is indicated by the formation of a clear zone. This research was conducted using extracts of Moringa oleifera leaves and Opuntia cochenillifera cladode extracted using ethyl acetate as a solvent. Antibiotic activity test against Staphylococcus aureus bacteria was done in Mannitol Salt Agar and Luria Bertani Agar media. The Kirby-Bauer discs used are Antimicrobial Disc Oxoid® and Filter Paper. The results of this study show that the use of the Antimicrobial Disc Oxoid® and Filter Paper shows no statistical difference in clear zone results (t-value = 0,45; p-value = 0,655; with α = 95%). While the Clear Zone results on Mannitol Salt Agar gave better results than Luria Bertani Agar and were statistically significant (t-value = 2,46; p-value = 0,02; with α = 95%). These findings indicate that filter paper can be an inexpensive alternative for the antibiotic test with no significantly different result compared to commercial antibiotic disc. However, an antibiotic test against Staphylococcus aureus is better done on Mannitol Salt Agar (MSA) compared to Luria Bertani Agar (LBA).
Co-Authors Adyan Donastin Endry Nugroho Prasetyo Endry Nugroho Prasetyo Maharani Pertiwi Koentjoro Maharani Pertiwi Koentjoro Setyo Nugroho