Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification

Four types of DNA extraction method from commercial dairy products like butter, chocolate and cheese were compared to identify the best DNA yield extractor. Among the four extraction method, the efficiency of CTAB method in extracting DNA was studied in detail with some modification including mechan...

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Main Authors: Rosman, Nurul Natasha, Mustafa, Shuhaimi, Khairil Mokhtar, Nur Fadhilah, Ali, Md. Eaqub
Format: Conference or Workshop Item
Language:English
Published: Halal Products Research Institute, Universiti Putra Malaysia 2014
Online Access:http://psasir.upm.edu.my/id/eprint/60520/1/M16.pdf
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author Rosman, Nurul Natasha
Mustafa, Shuhaimi
Khairil Mokhtar, Nur Fadhilah
Ali, Md. Eaqub
author_facet Rosman, Nurul Natasha
Mustafa, Shuhaimi
Khairil Mokhtar, Nur Fadhilah
Ali, Md. Eaqub
author_sort Rosman, Nurul Natasha
collection UPM
description Four types of DNA extraction method from commercial dairy products like butter, chocolate and cheese were compared to identify the best DNA yield extractor. Among the four extraction method, the efficiency of CTAB method in extracting DNA was studied in detail with some modification including mechanical lysis, manipulating amount of starting material for DNA extraction and testing different incubation temperature during DNA precipitation. The outcome presented here show that some of this modification have a significant effect on quality and quantity of extracted DNA from the dairy products. This modification allows for significantly higher yield of DNA from processed products than previously reported. The extracted DNA was than further evaluated using real-time PCR assay by targeting mitochondrial (mt) 16S ribosomal RNA (rRNA) gene and positively, all samples was amplifiable by this method. Furthermore, the swine-specific real-time PCR assay also can be performed on commercial samples targeting for 89 base pairs (bp) NADH dehydrogenase subunit 5 (ND5) gene from pork species for identification of pork derivatives in dairy products. These two methods were potentially reliable for food analysis and halal authentication.
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spelling upm.eprints-605202018-06-07T08:32:54Z http://psasir.upm.edu.my/id/eprint/60520/ Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification Rosman, Nurul Natasha Mustafa, Shuhaimi Khairil Mokhtar, Nur Fadhilah Ali, Md. Eaqub Four types of DNA extraction method from commercial dairy products like butter, chocolate and cheese were compared to identify the best DNA yield extractor. Among the four extraction method, the efficiency of CTAB method in extracting DNA was studied in detail with some modification including mechanical lysis, manipulating amount of starting material for DNA extraction and testing different incubation temperature during DNA precipitation. The outcome presented here show that some of this modification have a significant effect on quality and quantity of extracted DNA from the dairy products. This modification allows for significantly higher yield of DNA from processed products than previously reported. The extracted DNA was than further evaluated using real-time PCR assay by targeting mitochondrial (mt) 16S ribosomal RNA (rRNA) gene and positively, all samples was amplifiable by this method. Furthermore, the swine-specific real-time PCR assay also can be performed on commercial samples targeting for 89 base pairs (bp) NADH dehydrogenase subunit 5 (ND5) gene from pork species for identification of pork derivatives in dairy products. These two methods were potentially reliable for food analysis and halal authentication. Halal Products Research Institute, Universiti Putra Malaysia 2014 Conference or Workshop Item PeerReviewed text en http://psasir.upm.edu.my/id/eprint/60520/1/M16.pdf Rosman, Nurul Natasha and Mustafa, Shuhaimi and Khairil Mokhtar, Nur Fadhilah and Ali, Md. Eaqub (2014) Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification. In: Malaysia International Halal Research & Education Conference 2014 (MIHREC 2014), 2-4 Dec. 2014, Marriott Putrajaya Hotel, Malaysia. (pp. 194-201).
spellingShingle Rosman, Nurul Natasha
Mustafa, Shuhaimi
Khairil Mokhtar, Nur Fadhilah
Ali, Md. Eaqub
Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title_full Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title_fullStr Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title_full_unstemmed Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title_short Method optimizing DNA extraction on dairy products by real-time PCR targeting porcine-specific mitochondrial DNA gene for halal verification
title_sort method optimizing dna extraction on dairy products by real time pcr targeting porcine specific mitochondrial dna gene for halal verification
url http://psasir.upm.edu.my/id/eprint/60520/1/M16.pdf
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