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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.
EFEKTIVITAS KONSENTRASI DAN WAKTU MASERASI EKSTRAK DAUN BELUNTAS (Pluchea indica L.) TERHADAP PERTUMBUHAN Streptococcus pyogenes Ismi Yulandari, Siti; Dermawan, Asep; Kurniati, Iis; Iin Nur Indra, Asep
Jurnal Kesehatan Siliwangi Vol. 5 No. 2 (2024): JURNAL KESEHATAN SILIWANGI
Publisher : Politeknik Kesehatan Kemenkes Bandung

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Abstract

One of the most common diseases in Indonesia, namely pharyngitis, is caused by Streptococcus pyogenes. Inappropriate treatment of pharyngitis with antibiotics can result in bacteria that are resistant to antibacterials. Therefore, herbal plants that are effective as natural antibacterials are needed, such as beluntas leaves (Pluchea indica L.) to prevent this. Apart from that, the right method is needed so that the contents of the beluntas leaves can be attracted properly. The aim of this research was to determine the effective concentration and maceration time of beluntas leaf extract in inhibiting the growth of S.pyogenes. This research used varying maceration times of 24 and 72 hours. Then, from each variation of maceration time, beluntas leaf extract was made in concentrations of 5%, 10%, 15%, 20%, and 25%.  Beluntas leaf extract was tested for its inhibitory power against S.pyogenes using the Kirby Bauer method. The data obtained was the diameter of the inhibitory power of beluntas leaf extract on the growth of S.pyogenes, then the data was processed statistically using the Kruskal-Wallis test and a further test, namely the Post Hoc Test. The results of this research were that beluntas leaf extract which was macerated for 72 hours with a concentration of 20% and 25% had an average diameter of inhibition against S.pyogenes of 9.04 mm and 12.71 mm. Therefore, a maceration time of 72 hours with a concentration of 25% is effective in inhibiting the growth of S.pyogenes.
OPTIMASI VARIASI VOLTASE DAN WAKTU TERHADAP KUALITAS PITA DNA ESCHERICHIA COLI PADA PROSES ELEKTROFORESIS GEL AGAROSA nur amani putri, afifah; Iin Nur Indra, Asep; Merdekawati, Fusvita; Khoirul Abror, Yogi
Jurnal Kesehatan Siliwangi Vol. 5 No. 2 (2024): JURNAL KESEHATAN SILIWANGI
Publisher : Politeknik Kesehatan Kemenkes Bandung

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Abstract

Background: Escherichia coli is one of the causes of foodborne illness. Conventional PCR is a PCR method that is carried out qualitatively followed by visualization on agar electrophoresis. Agarose gel electrophoresis is a technique that is often used in various fields of science to separate a mixture of DNA on agarose substrate. This method is used to perform qualitative analysis of DNA samples. In electrophoresis, there are factors that affect the movement of DNA molecules, one of which is voltage. In addition, the length of time of the electrophoresis process can also affect the effectiveness of the results and the rate of migration. When the electrophoresis process time is short, large DNA fragments still tend to stick. Purpose: Therefore, it is necessary to balance the voltage and time given to get good DNA banding results. Methods: The research unit that will be used is the result of amplification of Escherichia coli 16SrRNA gene DNA measuring 584 bp. The electrophoresis process was carried out with voltage variations of 50, 100, 150 volts and time variations of 30, 45, 60 minutes. Observation of electrophoresis results that have formed DNA bands will be measured the area of DNA bands using ImageJ application. Conclusion: Based on the results of the study concluded that: The optimum voltage in the electrophoresis process in obtaining good E.coli DNA bands is at a voltage of 150 volts. The optimum time in the electrophoresis process in obtaining good E.coli DNA bands is for 30 minutes.
Pengembangan Marker Multiepitop untuk Kit Diagnostik HPV E6 Berbasis Lateral Flow Assay melalui Pendekatan In Silico Hasna, Nadifah; Rizki Febriyanti, Sabrina; Alginta, Taniani; Syifa Aprilia, Shafira; Fitri Sajidah, Ghaisani; Salsabila; Iin Nur Indra, Asep
JURNAL KESEHATAN KARTIKA Vol. 20 No. 2 (2025)
Publisher : Faculty of Health Science and Technology, University of Jenderal Achmad Yani

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26874/jkkes.v20i2.441

Abstract

Background : Cervical cancer is the second most common cancer after breast cancer and has a high mortality rate from all cancer deaths. Cervical cancer has been proven to be caused by continuous infection of oncogenic types of Human papillomavirus (HPV), namely high-risk HPV types including types 31, 33, 35, and 58. In the HPV life cycle, the E6 protein plays a crucial role in regulating cellular functions that contribute to cell transformation into cancer. The E6 protein from HPV types functions as the main oncoprotein that plays a role in the process of immortalization and cell malignancy. Mechanistically, the E6 protein from high-risk HPV contributes to the inactivation of the p53 tumor suppressor protein. Objective : This study aims to develop a multiepitope marker based on the E6 protein as the main component in the Lateral Flow Assay (LFA. Method : diagnostic kit with a bioinformatics approach method to identify immunogenic and conservative B cell epitopes in the HPV E6 protein which are then cloned in silico in the pET-28a (+) plasmid, so that it can produce a diagnostic kit that is specific for detecting Human papillomavirus (HPV). Result : The results showed that the designed multiepitope marker has the ability to trigger an effective immune response against B cells and has a fairly strong interaction between antigen and antibody. This study also showed the ability of the E6 protein to be expressed on the pET-28a(+) plasmid. Conclusion : This study provides a strong basis for further development and evaluation in vivo studies for the development of lateral flow assay (LFA)-based screening diagnostic kits utilizing Recombinant DNA technology.