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Journal : Science and Technology Indonesia

Assessment of Commercial DNA Extraction Kits for Porcine Gelatin Detection Using RT-PCR and ddPCR Gina, Safira; Rahmagiarti, Cintera; Ummah, Isna Mustafiatul; Sumantri, Cece; Suparto, Irma Herawati; Darmawan, Noviyan
Science and Technology Indonesia Vol. 9 No. 3 (2024): July
Publisher : Research Center of Inorganic Materials and Coordination Complexes, FMIPA Universitas Sriwijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26554/sti.2024.9.3.605-612

Abstract

Gelatin is extensively applied in various industries, including food, beverages, cosmetics, and pharmaceuticals. Although the determination of gelatin species is essential for religious, health, and consumer preference reasons, a standardized analysis method is absent. The challenge in identifying gelatin through DNA-based methods arises from the low DNA content and extensive DNA denaturation in the gelatin matrix. This study assessed the efficacy of two commercial DNA extraction kits, namely the Processed Food DNA Extraction (PF kit) and the DNeasy Mericon Food Kit (DM kit), for extracting DNA from porcine gelatin powder and commercial products derived from gelatin. Additionally, we evaluate the amplification of the extracted porcine DNA using real time polymerase chain reaction (RT-PCR) and droplet digital polymerase chain reaction (ddPCR) techniques. The PF extraction kit demonstrated successful DNA extraction from porcine gelatin powder and commercial samples of porcine gelatin-based candies with a higher concentration (32.24-286.07 ng/uL) and purity (A260/A280 ratio of 1.82-2.33) compared to the DM kit (3.95-7.30 ng/uL and an A260/A280 ratio of 1.29-2.45). RT-PCR and ddPCR analyses yielded positive results for porcine DNA from gelatin powder for both PF and DM kits, albeit with differing Cq values and copy numbers. The choice of DNA extraction kit significantly impacted the amplification results when analyzing commercial samples of porcine gelatin-based candies. Using RT-PCR, all samples yielded negative results with the DM kit, while the PF kit detected one positive result for porcine DNA. Improved outcomes were observed with more sensitive analysis methods such as ddPCR, where the DM kit identified one positive result for porcine DNA while the PF kit detected positive results for all tested candies.
Co-Authors . Hidayati . Subandriyo A Anggraeni A Anggraeni A B Dharmayanthi A Gunawan A M Fuah A.B.L. Ishak A.B.L. Ishak Abdul Samad ACHMAD FARAJALLAH Achmad Gitaraka Archimilar Adi Teruna Effendi Agus Buono Ahmad Furqon Ahmad Furqon Ahmad Furqon Ahmad Romadhoni Surya Putra ahmad yani Akbanugra Yudhananda Al Mukhlas Fikri Aldri Frinaldi Amalia Rahma Aminuyati Amri, Fisma Andi Baso Lompengeng Ishak Anggraeni Anggraeni Anggraeni, A. Anggraeni Ani Rosmayanti Anneke Anggraeni Anneke Anggraeni Anneke Anggraeni Anneke Anggraeni Any Aryani Arief Boediono Aron Batubara Asep Anang Asep Gunawan ASEP SAEFUDDIN Asmarasari, Santiananda Arta Astira, Adinda Wanda B Purwantara Bambang Purwantara Ben J. Takaendengan Ben J. Takaendengan Bramada Winiar Putra Budiariati, Vista Cahyo Budiman Cesilia M Dwiriani Chalid Talib Chalid Talib Clara Meliyanti Kusharto D D Solihin D, Nurhayati Dadan Rohdiana Dairoh Dairoh Dairoh Dairoh, Dairoh Darlian Fachruddin Dedi Duryadi Solihin Dedi Rahmat Depison Depison Depison Depison DEWI APRI ASTUTI Diah Nugrahani Pristihadi Dudi Syadili Dwi Budiono Dwi Margi Suci E Andreas E Taufik E. Andreas E. Andreas E. Damayanthi E. Damayanthi E. I. Rianti E. I. Rianti, E. I. Eko Handiwirawan Eko Handiwirawan Elmy mariana Elmy Mariana Epi Taufik Eryk Andreas Evy Damayanthi Fadhilah Dhani Santika Falah Farajallah, Ahmad Ferdian Achmad Gilang Kurnia Abdillah Gina, Safira Güler, Serdar Gushairiyanto Gushairiyanto Hadi Riyadi Harahap, Ahmad Saleh HARIMURTI MARTOJO Harini Nurcahya Mariandayani Hasrul Bani Asharudin Heni Rizqiati Heni Rizqiati Henny Nuraini herdis herdis Herpina Herpina Hidayat, Numan I Khaerunnisa I Wayan Mathius I wayan Teguh Wibawan Iman Supriatna Imas Batubara Irma . Irma . Irma Herawati Suparto Irma Irma Irma Irma Irma Isnafia Arief Irmanida Batubara Ishak, Andi Baso Lompengeng Ismeth Inounu Ismeth Inounu Isyana Khaerunnisa Isyana Khaerunnisa Iwan Prihantoro Jakaria Jakaria Jakaria Jakaria Jarmuji Jarmuji Jessy Mainidar Johar Arifin Joni Setiawan Junaedi Junaedi Junaedi Junaedi K Kinoshita Katrin Roosita Khaeruddin Khaeruddin Khaeruddin Khaeruddin Komang Alit Paramitasari Komariah . Kuswandi, Wawan Laelatul Choiriyah Lailatul Rohmah Lia Budimulyati Salman Lilis Nuraida Listyarini, Kasita Luci Cyrilla M Imron M S Engel M. Restu Mala Nurilmala Maskur Maskur Masrurah Masrurah Melly Pratiwi Setyawati Melly Pratiwi Setyawati Mochammad Sriduresta Soenarno Mohamad Hasil Tamzil Mohamad Yamin Mokhamad Fahrudin Mokhamad Fakhrul Ulum, Mokhamad Muh. Rusdin MUHAMMAD AGIL Muhammad Alwi Muhammad Fikri Al Habib Muhammad Suhendra, Muhammad Muhsinin, Muhammad Mutasem Ali M. Abuzahra N Hamashima N Hamashima Nena Hilmia Nensy Tri Putri Ni Wayan Kurniani Karja Niken Ulupi Nila Pratiwi Niswati, Za'imatun Noer Muhammad Dliyaul Haq Noraimah Binti Sulaiman Novita Nur Anggraeni Noviyan Darmawan Nurhayati D Nurhayati, Nurhayati D. Nu’man Hidayat Okti Nadia Poetri Olfa Mega Oloan Parlindungan Pardede, Berlin Pandapotan Pramujo, Muhammad Pratama, Bondan Purwantiningsih Sugita Puspita, Poppy Satya Putri Indah Ningtias R Diyono R Priyanto R. Iis Arifiantini Rahmagiarti, Cintera Rarah Ratih Adjie Maheswari Ratna Sholatia Harahap Ratnawati, Diana Raudah Putri Ekawati Refdanita, Refdanita Restu Misrianti Retno Setyaningsih Rina Martini Rini Herlina Mulyono Rita Mutia Romantis, Sara Rona Saumy Safitry Ronny R. Noor Ronny R. Noor Ronny Rachman Noor Ronny Rahman Noor RR Noor Rudi Afnan Rudi Priyanto Rudi Priyanto Rudy Priyanto Rukmiasih Rukmiasih S Darwati S Kahono S Sopiyana Saiful Anwar Salatnaya, H. Salsabila Ma’shum Imawan Santi Ananda Asmarasari Santiananda Arta Asmarasari Santoso Santoso Sarah Tazkya Sari Suryanah SATRIYAS ILYAS Septaria Jodiansyah Shidiq, Fajrin Sigid Prabowo Sikin Sikin Simon Elieser Siti Darodjah Siti Darodjah Rasad, Siti Siti Zubaidah Slamet Mulyanto Sony Hartono Wijaya Sri Adiani Sri Adiani Sri Darwati Sri Darwati Sri Mulatsih sri murtini . Sri Nurdiati SRI RAHAYU Sri Sulandari Sri Wahyuni Siswanti Sri Wahyuni Siswanti Subandriyo . . Sugyono . Sumiati Sumiati Sutikno Sutikno Sutikno Sutikno Sutikno Sutikno Syah, Setiawan Putra Syamsul Arifin T Akiyama T Morzumi T. Suryati TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Tatan Kostaman TIKE SARTIKA Tike Sartika Triana Susanti Triana Susanti Tuti Suryati Tuti Suryati Tuty Laswardi Yusuf Ummah, Isna Mustafiatul Vianisa, Putri Wasmen Manalu Weni Kurniati Widiatmaka Widya Pintaka Bayu Putra Wike andre Septian Winni Liani Daulay Yong Soo Kim Yulianto Yulianto Yun Alwi Yureni Sahril Dedi Yusman Setiawan Yusuf, Salman Zakiah Wulandari Zuraida Hanum Zuraida Hanum Zuraida Hanum