Nurul Faridah, - (2022) PEMANFAATAN PENANDA GENETIK DAN MULTIPLEX-PCR UNTUK DETEKSI CEPAT KEHALALAN PRODUK OLAHAN DAGING. S1 thesis, Universitas Pendidikan Indonesia.
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Abstract
Peningkatan konsumsi produk olahan daging menyebabkan peningkatan kasus pemalsuan produk olahan daging menggunakan daging tidak halal seperti babi dan tikus. Penggunaan penanda DNA dan Multiplex-PCR terus dikembangkan sebagai metode deteksi spesies pada produk olahan daging. Tujuan utama penelitian ini yaitu mendesain urutan primer multipleks babi, tikus, sapi, dan ayam yang menghasilkan amplikon yang dapat terbedakan melalui elektroforesis gel agarose untuk deteksi kehalalan spesies produk olahan daging. Desain primer dilakukan menggunakan data genom mitokondria pada situs NCBI-Primer BLAST untuk mendapatkan urutan primer babi dan sapi yang spesifik. Simulasi Single dan Multiplex-PCR secara in silico dilakukan menggunakan Primer Pooler. Validasi in vitro dilakukan melalui SinglePCR dan optimasi suhu annealing Multiplex-PCR menggunakan sampel daging ayam, sapi, babi, tikus dan produk olahan daging seperti bakso, sosis dan nugget. Hasil desain primer menemukan urutan primer babi dan sapi yang spesifik dengan gen target ND5 dan ukuran amplikon babi 860 bp dan sapi 1187 bp sedangkan hasil simulasi in silico menunjukkan bahwa primer multipleks berhasil ditemukan di dalam genom mitokondria dan mengamplifikasi sekuen target sehingga dihasilkan amplikon dengan ukuran terpanjang 1202 bp. Hasil validasi in vitro menemukan bahwa primer multipleks gen penanda halal berhasil mengamplifikasi seluruh sekuen target pada suhu annealing yang optimal yaitu 58oC dan hasil visualisasi elektroforesis gel agarose menunjukkan amplikon sapi (1187 bp), babi (860 bp), tikus (622 bp), dan ayam (272 bp) yang dapat terbedakan. Amplifikasi primer multipleks gen penanda halal menggunakan sampel produk olahan daging dapat mendeteksi spesies yang digunakan pada produk olahan daging sehingga kehalalannya dapat diketahui. Kata kunci: Deteksi kehalalan, Desain primer, Multiplex-PCR, Penanda DNA, Produk Olahan Daging The increase in consumption of processed meat products has led to an increase in cases of adulterating processed meat products using non-halal meat such as pork and rats. The use of DNA markers and Multiplex-PCR continues to be developed as a method of species detection in processed meat products. The main objective of this study was to design a sequence of multiplex primers for pigs, rats, cattle, and chickens that produced distinguishable amplikon through agarose gel electrophoresis to detect the halal species of processed meat products. The primer design was performed using mitochondrial genom data at the NCBI-Primary BLAST site to obtain specific pig and bovine primer sequences. In silico Single and Multiplex-PCR simulations were performed using the Primer Pooler. In vitro validation was carried out through SinglePCR and multiplex-PCR annealing temperature optimization using samples of chicken, beef, pork, rat and processed meat products such as meatballs, sausages and nuggets. The results of the primary design of specific pig and bovine primary sequences with the target gene ND5 and the size of the amplikon in pigs 860 bp and cattle 1187 bp, while the in silico simulation results show that the primary multiplex was found in the mitochondrial genom and amplified the target sequence to produce the amplikons with the longest size. 1202 bp. The results of in vitro validation found that the multiplex primers for the halal marker gene were successful in amplifying all target sequences at the optimal annealing temperature of 58oC and the agarose gel electrophoresis visualization showed amplikons of cattle (1187 bp), pigs (860 bp), rat (622 bp), and chicken (272 bp) that could be distinguished. The multiplex primer amplification of halal marker genes using samples of processed meat products can detect the species used in processed meat products so that halalness can be determined. Keywords: DNA markers, Halal detection, Multiplex-PCR, Primer design, Processed Meat Products
| Item Type: | Thesis (S1) |
|---|---|
| Additional Information: | https://scholar.google.com/citations?hl=id&user=0QCf3MsAAAAJ ID Sinta Dosen Pembimbing Diah Kusumawaty: 6005459 Didik Priyandoko: 6658318 |
| Uncontrolled Keywords: | Deteksi kehalalan, Desain primer, Multiplex-PCR, Penanda DNA, Produk Olahan Daging |
| Subjects: | L Education > L Education (General) Q Science > QH Natural history > QH301 Biology |
| Divisions: | Fakultas Pendidikan Matematika dan Ilmu Pengetahuan Alam > Jurusan Pendidikan Biologi > Program Studi Biologi (non kependidikan) |
| Depositing User: | Nurul Faridah |
| Date Deposited: | 08 Sep 2022 03:41 |
| Last Modified: | 08 Sep 2022 03:41 |
| URI: | http://repository.upi.edu/id/eprint/78991 |
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