Putri, Sekar Ramadhanti and Rahma Micho Widyanto, S.Si, MP. and Dr. Irma Sarita Rahmawati, STP., MP., MSc. (2021) Optimasi Metode Analisis Daging Babi Berbasis DNA Menggunakan Loop-Mediated Isothermal Amplification dengan Pewarna Diamond Nucleic Acid dan Calcein. Sarjana thesis, Universitas Brawijaya.
Abstract
Daging merupakan bahan makanan yang kaya akan zat gizi, namun daging merah masih sulit dibedakan secara fisik oleh masyarakat. Sedangkan banyak terjadi kasus adulterasi daging mentah dengan daging babi di Indonesia. Sehingga diperlukan metode pengujian untuk menjamin kehalalan bahan makanan dengan mendeteksi adanya DNA babi pada bahan makanan secara sederhana dan spesifik menggunakan Loop-Mediated Isothermal Amplification (LAMP) dan divisualisasikan dengan pewarna, dua diantaranya yaitu Diamond Nucleic Acid dan Calcein. Penelitian ini bertujuan untuk mengidentifikasi perbedaan hasil optimasi analisis daging babi menggunakan metode LAMP dengan jenis pewarna Diamond Nucleic Acid dan Calcein. Desain penelitian yaitu true experimental laboratory dengan perlakuan berupa penambahan pewarna Diamond Nucleic Acid dan Calcein pada sampel Nuclease free H2O, DNA sapi dan DNA babi menggunakan uji LAMP. DNA sapi dan babi didapatkan dari proses isolasi DNA dan pengukuran nanodrop. Hasil penelitian menunjukkan rasio kemurnian DNA babi sebesar 1,76 dan 1,82 serta DNA sapi sebesar 1,89 dan 1,92. Dengan hasil visualisasi menggunakan Diamond dye pada percobaan 1 yaitu berpendar pada tube DNA babi dan tidak berpendar pada tube kontrol dan DNA sapi. Serta hasil percobaan kedua dan ketiga yaitu berpendar pada seluruh tube. Sedangkan hasil visualisasi menggunakan Calcein percobaan pertama dan kedua menunjukkan hasil berpendar pada seluruh tube. Kesimpulan dari penelitian ini adalah pewarna Diamond dan Calcein mampu menunjukkan hasil LAMP dengan adanya perubahan warna yang terjadi namun kedua pewarna memiliki perbedaan dari segi mekanisme kerja, metode penambahan pewarna, keamanan dan sensitivitas.
English Abstract
Meat is one of food that rich in nutrients, but it is difficult to physically distinguish between red meat. Meanwhile, there are many cases adulteration of raw meat with pork in Indonesia. Therefore, Loop-Mediated Isothermal Amplification (LAMP) is one of simple and specific method that needed to ensure the food is halal by detecting the presence of pork DNA in food sample. This method can be visualized by adding dye, two of dye that can be used is Diamond Nucleic Acid and Calcein. This study aims to identify differences in optimization results of pork analysis using the LAMP method with different kind of dye, Diamond Nucleic Acid and Calcein. True experiment laboratory conducted with the addition of Diamond Nucleic Acid dyes and Calcein to three different samples that is Nuclease free H2O, DNA from beef and pork using LAMP method. DNA from beef and pork obtained from the isolation process and measured by nanodrop. The result of this study showed that the purity of DNA used in this experiment is 1,76 and 1,82 for pork as well for beef is 1,89 and 1,92. With the result of visualization using Diamond dye in first experiment, showed fluorescent on the pork tube and not fluorescent on the control and beef tube. Furthermore, the results of the second and third experiments were fluorescent on all tubes. Meanwhile, the results of visualization using Calcein in the first and second experiments showed fluorescent results in all tubes. The conclusion of this study is that both dye are able to visualize LAMP results with color change that occurs after reaction, but Diamond and Calcein have difference in terms of their mechanism of action, method when adding dye, safety and sensitivity.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 0521060187 |
| Uncontrolled Keywords: | Adulterasi, loop mediated isothermal amplification (LAMP), diamond nucleic acid, calcein, daging babi, Adulteration, loop mediated isothermal amplification (LAMP), diamond nucleic acid, calcein, pork |
| Subjects: | 600 Technology (Applied sciences) > 613 Personal health and safety |
| Divisions: | Fakultas Kedokteran > Ilmu Gizi |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 22 Dec 2021 06:23 |
| Last Modified: | 24 Feb 2022 04:44 |
| URI: | http://repository.ub.ac.id/id/eprint/187592 |
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Text (DALAM MASA EMBARGO)
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