VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR)
Porcine contaminant on nugget have been analyzed using real-time Polymerase Chain Reaction (RTi-PCR). The aim of this study is to develop porcine identification method on nugget based DNA which can be used as halal authentication. Real-time PCR amplified ND5 target gene in pork�s mithochondria DNA...
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| Format: | Thesis |
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[Yogyakarta] : Universitas Gadjah Mada
2013
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| author | , IMELDA O. TAMPUBOLON , Dr. Tri Joko Raharjo, M.Si |
| author_facet | , IMELDA O. TAMPUBOLON , Dr. Tri Joko Raharjo, M.Si |
| author_sort | , IMELDA O. TAMPUBOLON |
| collection | UGM |
| description | Porcine contaminant on nugget have been analyzed using real-time Polymerase Chain Reaction (RTi-PCR). The aim of this study is to develop porcine identification method on nugget based DNA which can be used as halal authentication.
Real-time PCR amplified ND5 target gene in pork�s mithochondria DNA using species-specific primer, namely forward primer ND5 (5�-CATTCGCCTCACTCACATTAACC-3�), and reverse primer ND5 (5�-AAGAGAGAGTTCTACGGTCTGTAG-3�). The steps of this study consisted of determining the annealing temperature (Ta) and the method validation include the specificity, precision and determination of the limit detection. Real-time PCR application used for detection porcine contamination on nuggets. DNA was isolated using phenol KIAA. Amplification of DNA fragments was conducted with PCR conditions : pre-denaturation 95 °C for 30 s, denaturation 95 °C for 2 seconds and annealing at optimal Ta result for 5 seconds.
DNA amplification using real-time PCR with primers ND5 produced an amplicon with a melting point of 79 °C and 227 base pair fragment through agarose gel electrophoresis. The optimal annealing temperature was 58 °C. Validated method had high specificity and sensitivity, specific primers amplified DNA ND5 pigs and the detection limit up to 1% porcine contamination in food products. Seven samples were tested and proved not contain pork. |
| first_indexed | 2024-03-13T22:57:08Z |
| format | Thesis |
| id | oai:generic.eprints.org:121756 |
| institution | Universiti Gadjah Mada |
| last_indexed | 2024-03-13T22:57:08Z |
| publishDate | 2013 |
| publisher | [Yogyakarta] : Universitas Gadjah Mada |
| record_format | dspace |
| spelling | oai:generic.eprints.org:1217562016-03-04T08:38:57Z https://repository.ugm.ac.id/121756/ VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) , IMELDA O. TAMPUBOLON , Dr. Tri Joko Raharjo, M.Si ETD Porcine contaminant on nugget have been analyzed using real-time Polymerase Chain Reaction (RTi-PCR). The aim of this study is to develop porcine identification method on nugget based DNA which can be used as halal authentication. Real-time PCR amplified ND5 target gene in pork�s mithochondria DNA using species-specific primer, namely forward primer ND5 (5�-CATTCGCCTCACTCACATTAACC-3�), and reverse primer ND5 (5�-AAGAGAGAGTTCTACGGTCTGTAG-3�). The steps of this study consisted of determining the annealing temperature (Ta) and the method validation include the specificity, precision and determination of the limit detection. Real-time PCR application used for detection porcine contamination on nuggets. DNA was isolated using phenol KIAA. Amplification of DNA fragments was conducted with PCR conditions : pre-denaturation 95 °C for 30 s, denaturation 95 °C for 2 seconds and annealing at optimal Ta result for 5 seconds. DNA amplification using real-time PCR with primers ND5 produced an amplicon with a melting point of 79 °C and 227 base pair fragment through agarose gel electrophoresis. The optimal annealing temperature was 58 °C. Validated method had high specificity and sensitivity, specific primers amplified DNA ND5 pigs and the detection limit up to 1% porcine contamination in food products. Seven samples were tested and proved not contain pork. [Yogyakarta] : Universitas Gadjah Mada 2013 Thesis NonPeerReviewed , IMELDA O. TAMPUBOLON and , Dr. Tri Joko Raharjo, M.Si (2013) VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR). UNSPECIFIED thesis, UNSPECIFIED. http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=61853 |
| spellingShingle | ETD , IMELDA O. TAMPUBOLON , Dr. Tri Joko Raharjo, M.Si VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title | VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title_full | VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title_fullStr | VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title_full_unstemmed | VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title_short | VALIDASI UJI KONTAMINASI BABI DALAM NUGGET DENGAN METODE REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR) |
| title_sort | validasi uji kontaminasi babi dalam nugget dengan metode real time polymerase chain reaction rti pcr |
| topic | ETD |
| work_keys_str_mv | AT imeldaotampubolon validasiujikontaminasibabidalamnuggetdenganmetoderealtimepolymerasechainreactionrtipcr AT drtrijokoraharjomsi validasiujikontaminasibabidalamnuggetdenganmetoderealtimepolymerasechainreactionrtipcr |