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All Journal Biogenesis: Jurnal Ilmiah Biologi Science and Technology Indonesia
Ummah, Isna Mustafiatul
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Environmental Science Immunology & microbiology Materials Science & Nanotechnology Physics

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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.
Morphological Traits, Productive Performance and Genotyping Fat Deposition PPAR Gene in Gama Ayam Crossbreeds of Female F1 Kamper and Male BC1 Kambro Ummah, Isna Mustafiatul; Mahardhika, I Wayan Swarautama; Daryono, Budi Setiadi
Biogenesis: Jurnal Ilmiah Biologi Vol 7 No 2 (2019)
Publisher : Department of Biology, Faculty of Sci and Tech, Universitas Islam Negeri Alauddin Makassar

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24252/bio.v7i2.9993

Abstract

On the recent classification of native Indonesian chicken, there are 31 breeds of chicken. Highly circulated chicken breed in Indonesia is fast-growing broiler that includes several strains such as Cobb 500, Hubbard, Hybro, Isa Hyline, and Hisex. Fast-growing broiler chickens have rapid growth, with excessive fat deposition on chicken carcass which implicates a health problem and unfavorable meat quality. Gama Ayam Research Team conducted selective breeding which produces two new chicken breed Kamper and Kambro. Further selective breeding then crossbred female F1 Kamper and male BC1 (backcross I) Kambro. In this research morphological traits, productive performance and genotyping of PPAR gene related to fat deposition gene and blood lipid content in Gama Ayam were identified. Based on data analysis hybrid chickens morphological traits it concluded that Gama Ayam has a significant variation based on feather color. The productive performance was determined with the feed-conversion ratio (FCR) value which was 3.17. Genotyping of PPAR gene resulted in four polymorphisms that formed 14 haplotypes groups. Based on blood lipid content analysis of cholesterol content, triglycerides content, HDL and LDL, Gama Ayam have significantly lower content of cholesterol (107.05 mg/dl), LDL (44.71 mg/dl) and triglycerides (22.41 mg/dl). PPAR gene polymorphisms are not correlated with blood lipid content in Gama Ayam. A significantly strong correlation between PPAR gene polymorphism on the bodyweight of Gama Ayam at 49-days-old. Triglyceride level, cholesterol level and LDL level in Gama Ayam were lower than broiler chicken. Further research with a larger population size and sex classification of hybrid chicken must be conducted to validate the results.
Co-Authors BUDI SETIADI DARYONO Cece Sumantri Gina, Safira Irma Herawati Suparto Mahardhika, I Wayan Swarautama Noviyan Darmawan Rahmagiarti, Cintera