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All Journal Current Biomedicine Mukhtabar: Journal of Medical Laboratory Technology Jurnal Kesehatan Siliwangi
Djuminar, Ai
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Biochemistry, Genetics & Molecular Biology Chemistry Dentistry Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology Public Health Veterinary

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Konsentrasi dan kemurnian ekstraksi DNA metode sonikasi dan spin column dari sampel dahak penderita tuberkulosis Saputra, Fitrianingsih; Indra, Asep Iin Nur; Djuminar, Ai; Merdekawati, Fusvita; Nurhayati, Betty
Current Biomedicine Vol. 2 No. 2 (2024): July
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.2.2.84-92

Abstract

Background: The Polymerase Chain Reaction (PCR) method can identify Mycobacterium tuberculosis in a sputum sample of a patient with TB (TB). One crucial step to ensure accurate PCR results is the DNA extraction process. Objective: The research aims to compare the concentration and purity of DNA from the sputum of TB patients using ultrasound and spin column extraction techniques. Methods: The research uses descriptive study designs with post-only design strategies. The primary data was derived from 18 sputum specimens from TB patients. Concentration measurement and DNA purity testing using a nanodrop spectroscopic photometer. Results: DNA extraction by ultrasound method has an average concentration of 18.9 ± 8.5 ng/L, with a peak of 37.6 ng/ L. The spin column method produces an average of 55.5 ± 27.9 ng/μL; the peak is 105.0 ng/ μL. The purity value of the DNA extract is in the range of 1.8 ± 2.0 with the ultrasound method of 61% and the spin column of 78%. Conclusion: The sonication method has a lower average concentration and a higher percentage of purity than the spin column method, and there are differences in concentrations and purity values between the two methods.
OPTIMASI VARIASI KONSENTRASI AGAROSE DAN WAKTU ELEKTROFORESIS TERHADAP KUALITAS PITA DNA Mycobacterium tuberculosis RESISTAN ISONIAZID Halimah, Ina; Firman Solihat , Mohamad; Djuminar, Ai; Rismiati, Zuri
Jurnal Kesehatan Siliwangi Vol. 4 No. 3 (2024): JURNAL KESEHATAN SILIWANGI
Publisher : Politeknik Kesehatan Kemenkes Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.34011/jks.v4i3.1854

Abstract

Mycobacterium tuberculosis resistan isoniazid (INH) terjadi akibat mutasi gen dimana sebagian besar terjadi pada gen katG. Pada penelitian ini digunakan sampel dengan primer untuk mendeteksi mutasi gen katG untuk MRD TB pada kodon 315 dan 463 dengan panjang produk primer yang bervariasi diantaranya 101 bp (untuk mutasi S315T), 141 bp (mutasi S315N), 200 bp (mutasi S315I), 247 bp (mutasi S315R), 290 bp (mutasi S315G), dan 400 bp (mutasi R463L). Penelitian ini bertujuan untuk mengetahui konsentrasi dan lama waktu elektroforesis Mycobacterium tuberculosis resistan isoniazid (INH) pada gel agarose gel agarose yang optimum. Jenis penelitian yang digunakan adalah eksperimen mengenai perbandingan variasi konsentrasi agarose 1%, 1,5% dan 2% terhadap variasi lama waktu elektroforesis pada 30 menit, 45 menit dan 60 menit. Analisa pita DNA hasil elektroforesis menggunakan aplikasi imageJ. Pada penelitian ini sampel yang digunakan adalah pull DNA Mycobacterium tuberculosis resistan isoniazid hasil produk real time PCR. Hasil penelitian menunjukkan bahwa pita DNA Mycobacterium tuberculosis resistan isoniazid (INH) gen katG untuk MRD TB pada kodon 315 dengan ukuran produk 101 bp S315T, 141 bp S315N, 200 bp S315I, dan 290 bp S315G diperoleh optimal pada konsentrasi agarose 2% dan waktu elektroforesis selama 45 menit. Pita DNA Mycobacterium tuberculosis resistan isoniazid (INH) gen katG untuk MRD TB pada kodon 315 dengan ukuran produk 247 bp S315R dan gen katG untuk MRD TB pada kodon 463, ukuran 400 bp R463L optimal pada konsentrasi agarose 2% dengan waktu elektroforsis selama 60 menit.
Optimization of Concentration and Staining Duration of Methyl Green in The Examination of Escherichia coli DNA Bands Using Agarose Gel Electrophoresis Adrian Prasetya, Rifky; Merdekawati, Fusvita; Iin Nur Indra, Asep; Djuminar, Ai
Mukhtabar Journal of Medical Laboratory Technology Vol 2 No 2 (2024): Mukhtabar: Journal of Medical Laboratory Technology (October 2024)
Publisher : LPPM STIKes Muhammadiyah Ciamis

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52221/mjmlt.v2i2.696

Abstract

Background & Objective: Nowadays, many alternative dyes are used for staining DNA during electrophoresis, one of which is Methyl Green. Methyl Green has several advantages, including being cheaper than EtBr, having low toxicity, being non-carcinogenic, and as a cationic dye, Methyl Green is resistant to photobleaching. This study aims to determine the optimal concentration and staining duration of Methyl Green as a DNA dye for examining Escherichia coli DNA bands using agarose gel electrophoresis. Method: The research method used is experimental. Amplified Escherichia coli 16S rRNA gene DNA, sized 584 bp, which has undergone electrophoresis, was stained with Methyl Green dye at concentrations of 0.10%, 0.15%, 0.20%, 0.25%, 0.00015%, 0.00020%, 0.00025%, and 0.00030% with varying immersion times of 10, 15, 20, and 25 minutes. Result: The resulting DNA bands were analyzed or measured for surface area using ImageJ software. The mean value for each experimental group was calculated. The highest mean value was used as the basis for determining the most optimal condition. Conclusion: This study concludes that the optimal concentration and staining duration of Methyl Green, based on the highest mean value of 19,844,845, is 0.00030% Methyl Green with a staining duration of 25 minutes.
Co-Authors Adrian Prasetya, Rifky Firman Solihat , Mohamad Halimah, Ina Iin Nur Indra, Asep Indra, Asep Iin Nur Merdekawati, Fusvita Nurhayati, Betty Rismiati, Zuri Saputra, Fitrianingsih