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All Journal Pharmaciana: Jurnal Kefarmasian Biota INDONESIAN JOURNAL OF MEDICAL LABORATORY SCIENCE AND TECHNOLOGY Jurnal Pendidikan dan Konseling Jurnal Pembelajaran dan Biologi Nukleus Jurnal Ners Community Development Journal: Jurnal Pengabdian Masyarakat Jurnal Integrasi Kesehatan dan Sains Jurnal Ilmiah Wahana Pendidikan Journal of Pharmaceutical and Sciences. Jurnal Abdimas Mahakam
Jekmal Malau
Program Studi Farmasi Universitas Singaperbangsa Karawang, Indonesia
Religion Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Dentistry Economics, Econometrics & Finance Education Environmental Science Health Professions Immunology & microbiology Languange, Linguistic, Communication & Media Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Nursing Public Health Social Sciences Other

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Journal : biota

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Comparison of Two PCR Primer Sets for In-House Validation of GHSR Gene Variation Detection Employing Artificial Recombinant Plasmid Approach Ahsanal Kasasiah; Jekmal Malau; Sekar Andjung Tresnawati; Priscinya Christiana Debora; Nur Komala Fitri; Saarah Hamidah Asmara Indratno; Asman Hitopik; Eriyanti Astika; Anisa Aula Rahma; Fikri, Al Mukhlas
Biota Vol 10 No 2 (2024): Jurnal Biota 2024
Publisher : Faculty of Science and Technology Universitas Islam Negeri Raden Fatah Palembang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19109/biota.v10i2.21166

Abstract

Stunting is a significant global public health problem caused by long-term dietary deficits that affect many children worldwide. Both environmental and genetic factors, including variants in the GHSR gene, play a crucial role in stunted growth. This study used an artificial recombinant plasmid DNA method to evaluate two primer set combinations for identifying DNA variants in the GHSR gene. Selecting suitable primer sets for identifying GHSR genetic variants linked to stunting is essential, as evidenced by PCR and sequencing techniques. The target gene, based on the GHSR reference sequence, consists of eight DNA variations (ΔQ36, G57G, P108L, L118L, R159R, C173R, D246A, and A277P). A recombinant plasmid was created by inserting a 1000 bp fragment of the GHSR gene into the pUC57 backbone. Primer sets were chosen based on their capacity to amplify these eight genetic variations and were optimized and validated using PCR methods. PCR and bi-directional sequencing verified the existence of surrounding DNA and specific single nucleotide variants (SNVs). In our study, we discovered four changes in the DNA sequence (R159R G>A, C173R T>C, D246A A>C, and A277P G>C) using the E1_F2/E1_R3 primer pair. Additionally, a new combination of primers (E1_F1/E1_R3) effectively detected seven DNA sequence mutations (ΔQ36 del CAG, G57G C>T, P108L C>T, L118L C>T, R159R G>A, C173R T>C, and D246A A>C). We have developed a new combination of forward and reverse primers to identify seven SNVs in the GHSR gene, which could serve as a diagnostic tool in clinical laboratory settings.
Co-Authors Afif Tri Nugraha Ahsanal Kasasiah Al Mukhlas Fikri Alfina Oktavianti Aliya Azkia Zahra Aliza Salsabila Ainaputri Angel Novia Fransiska Anggi Ayu Pratama Anisa Aula Rahma Anisa Aula Rahma Anisa Fauziah Anjarwati, Sri Ari Purwanti Arline Rahmadianti Kusumawardhani Asman Hitopik Asman Hitopik Asri Yuniar Cantika Aprillia Chalisya Vanya Advaita Datu Muhammad Cordova Devi Ratnasari Dewi Pratiwi Purba Siboro Dhavina Maharani Diva Afiah Hanifa Irawan Eka Melati Elvira Julia Ariyanto Endeh Apriyanti Eriyanti Astika Eriyanti Astika Faizal Auladi Rivianto Fikri, Al Mukhlas Herdiana Verliani Indah Laily Hilmi Irene Virda Sakina Isyana S alsabila Lestari Mahardika Urbaningrum Mekar Meilisa Amalia Mochamad Galuh Ryandha Muhammad Fajri Maariz Mulidini Mulidini Munir Alinu Mulki Mutia Desvi Putri Mutiara Alfiyah Najla Yusiana Wahyudi Nissa Khalida Hanum Nofirman, Nofirman Nolla Olipia Elva Megrian Nur Komala Fitri Nurma Dwi Rahmawati Olivia Tahalele Priscinya Christiana Debora Priscinya Christiana Debora Putri Setya Tyasna Putri Wulanbirru Ramdhan Kurniawan Saarah Hamidah Asmara Indratno Saarah Hamidah Asmara Indratno Salsabila Granadha Nibullah Sekar Andjung Tresnawati Shafira Intan Anggraini Shinta Puspa Dwiyanti Siti Nafisa Siti Nuriah Siti Rohmah Sofianti Hidayat Sri Rahayu Syifa Khairunnisa Ti Aisyah Tiara Cinta Amelia Tiara Nurayuni Tintia Rafika Putri Tiwi Ambarati Vina Luthfiana Hasna Vriezka Mierza Welly Windari Weraman, Pius Yuliana Arsyad Yumareta Anggun Zuyyinna Alya Abbas