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All Journal Journal Of Vocational Health Studies Jurnal Kesehatan Siliwangi
Asep Iin Nur Indra
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Biochemistry, Genetics & Molecular Biology Dentistry Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology

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OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS Fajri, Siti Nuristiqomah; Asep Iin Nur Indra; Fusvita Merdekawati; H. , Acep Tantan
Journal of Vocational Health Studies Vol. 8 No. 1 (2024): July 2024 | JOURNAL OF VOCATIONAL HEALTH STUDIES
Publisher : Faculty of Vocational Studies, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jvhs.V8.I1.2024.35-41

Abstract

Background: Deoxyribonucleic acid (DNA) molecules can be effectively visualized when stained and observed under ultraviolet light during the electrophoresis process. The commonly used dye Ethidium bromide (EtBr) is considered hazardous due to its potential to cause mutations, cancer, and congenital disabilities. Various alternative dyes have been reported, one of which is hematoxylin. Hematoxylin compounds do not have mutagenic potential and are easier to apply than EtBr. However, there is no optimal variation in concentration and duration of staining for clear and effective visualization of DNA bands. Purpose: To find the concentration and staining time of harris hematoxylin staining for DNA bands from agarose gel electrophoresis. Method: An experimental method with a group comparison statistical design. The amplified DNA I6S rRNA gene from Escherichia coli (584 bp), which had undergone electrophoresis, was stained using harris hematoxylin dye at 0.01%; 0.02%; 0.03%; 0.04%; and 0.05% concentrations and immersion times soaking times of 10, 15, 20, and 30 minutes variations. The intensity of the DNA bands was analyzed using ImageJ. The staining power of the experimental groups was compared to the intensity of control dye and given a grading score of 1 - 4. The experiment was repeated twice, and the mean grading score was calculated. The highest mean value was considered the most optimal value. Result: A concentration of 0.02% showed relatively constant staining intensity for each soaking time. A mean value of 3.5 was obtained for a 0.01% concentration for 15 minutes. A 0.03% and 0.04% concentrations for 20 minutes. Conclusion: The highest mean value of 4 was obtained for Harris hematoxylin at 0.05% for 15 minutes.
Comparison of Color Quality in Tissue Preparations using Methyl Ester Sulfonate And Xylol as a Deparafinization Agent Rahmah, Syafa Aulia; Adang Durachim; Wiwin Wiryanti; Asep Iin Nur Indra
Jurnal Kesehatan Siliwangi Vol. 5 No. 2 (2024): JURNAL KESEHATAN SILIWANGI
Publisher : Politeknik Kesehatan Kemenkes Bandung

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Abstract

Deparaffinization is the process of decaying the remnants of paraffin in tissue preparations. Deparaffinization aims to make the dye that will be given to the preparation fully absorbed so that the preparation can be colored properly. This deparaffinization process uses xylol solution in its immersion. Methyl Ester Sulfonate (MES) is one of the vegetable oil-based anionic surfactants that can dissolve in water and has a long carbon chain that is soluble in oil and vaseline. The purpose of this study was to determine whether Methyl Ester Sulfonate can be used as a deparaffinizing agent for tissue preparations. The sample in this study was appendiceal tissue as many as 30 units, each unit of tissue made into 2 tissue preparations so that it became 60 tissue preparations, namely 30 tissues using xylol as a deparaffinizing agent and 30 tissues using methyl ester sulfonate. The parameters of this study were the clarity of the color of the nucleus and cytoplasm, the uniformity of cell color, and the contrast of the color of the nucleus and cytoplasm qualitatively and quantitatively. The statistical test used was Independent T-Test test if the data were normally distributed and Mann Whitney test if the data were not normally distributed. Based on the Independent T-Test test, the Sig value was obtained. 0.951 and 0.102, both of which are >0.05, meaning that there is no significant difference between the use of xylol and 10% methyl ester sulfonate as a deparaffinizing agent.
Co-Authors Adang Durachim Fajri, Siti Nuristiqomah Fusvita Merdekawati H. , Acep Tantan Rahmah, Syafa Aulia Wiwin Wiryanti