OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS
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...
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Universitas Airlangga
2024-07-01
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| Series: | Journal of Vocational Health Studies |
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| Online Access: | https://e-journal.unair.ac.id/JVHS/article/view/48061 |
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| author | Siti Nuristiqomah Fajri Asep Iin Nur Indra Fusvita Merdekawati Acep Tantan H. |
| author_facet | Siti Nuristiqomah Fajri Asep Iin Nur Indra Fusvita Merdekawati Acep Tantan H. |
| author_sort | Siti Nuristiqomah Fajri |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-dad3f939d3a14962ba1d37de6e622236 |
| institution | Kabale University |
| issn | 2580-7161 2580-717X |
| language | English |
| publishDate | 2024-07-01 |
| publisher | Universitas Airlangga |
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| series | Journal of Vocational Health Studies |
| spelling | doaj-art-dad3f939d3a14962ba1d37de6e6222362025-02-03T05:46:26ZengUniversitas AirlanggaJournal of Vocational Health Studies2580-71612580-717X2024-07-0181354110.20473/jvhs.V8.I1.2024.35-4146148OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESISSiti Nuristiqomah Fajri0https://orcid.org/0000-0002-9935-8111Asep Iin Nur Indra1https://orcid.org/0009-0005-9235-7932Fusvita Merdekawati2https://orcid.org/0009-0000-9293-9610Acep Tantan H.3https://orcid.org/0009-0000-3661-8783Medical Laboratory Technology, Bandung Ministry of Health Polytechnic, CimahiMedical Laboratory Technology, Bandung Ministry of Health Polytechnic, CimahiMedical Laboratory Technology, Bandung Ministry of Health Polytechnic, CimahiMedical Laboratory Technology, Bandung Ministry of Health Polytechnic, CimahiBackground: 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.https://e-journal.unair.ac.id/JVHS/article/view/48061electrophoresisconcentrationsoaking timeharris hematoxylin |
| spellingShingle | Siti Nuristiqomah Fajri Asep Iin Nur Indra Fusvita Merdekawati Acep Tantan H. OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS Journal of Vocational Health Studies electrophoresis concentration soaking time harris hematoxylin |
| title | OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS |
| title_full | OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS |
| title_fullStr | OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS |
| title_full_unstemmed | OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS |
| title_short | OPTIMIZATION OF CONCENTRATION AND SOAKING TIME OF HARRIS HEMATOXYLIN IN DNA BAND EXAMINATION USING ELECTROPHORESIS |
| title_sort | optimization of concentration and soaking time of harris hematoxylin in dna band examination using electrophoresis |
| topic | electrophoresis concentration soaking time harris hematoxylin |
| url | https://e-journal.unair.ac.id/JVHS/article/view/48061 |
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