Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis
Successful DNA amplification is vital for the detection of specific DNA targets in feeds, and this in return depends on the ability of DNA extraction methods to produce good quality DNA. In this study, seven methods were compared for DNA extraction from feeds using quantitative polymerase chain re...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Faculty of Food Science and Technology, Universiti Putra Malaysia
2009
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| Online Access: | http://psasir.upm.edu.my/id/eprint/16439/1/16439.pdf |
| _version_ | 1825945816374706176 |
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| author | Kesar Singh, Jasbeer Kaur Radu, Son Mohamad Ghazali, Farinazleen Cheah, Yoke Kqueen |
| author_facet | Kesar Singh, Jasbeer Kaur Radu, Son Mohamad Ghazali, Farinazleen Cheah, Yoke Kqueen |
| author_sort | Kesar Singh, Jasbeer Kaur |
| collection | UPM |
| description | Successful DNA amplification is vital for the detection of specific DNA targets in feeds, and this in return depends on the ability of DNA extraction methods to produce good
quality DNA. In this study, seven methods were compared for DNA extraction from feeds using quantitative polymerase chain reaction (PCR) of single copy maize (Zea mays) endogenous hmg (high mobility group) gene. Relative levels of hmg were used to evaluate the DNA quality.
Spectrophotometer determination of DNA was also carried out to assess DNA yield and DNA purity, while electrophoretic analysis of genomic DNA extracts was carried out to investigate DNA integrity. The findings illustrate that the DNA extraction methods have a significant effect on DNA quality. Statistically, the Epicentre method extracted the highest DNA yield while the Wizard method had the lowest DNA yield with high DNA purity and integrity. However, the
Wizard method recovered the most amplifiable DNA per reaction, indicating that template quality and integrity had greater influence over hmg amplification than DNA yield. |
| first_indexed | 2024-03-06T07:37:26Z |
| format | Article |
| id | upm.eprints-16439 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:37:26Z |
| publishDate | 2009 |
| publisher | Faculty of Food Science and Technology, Universiti Putra Malaysia |
| record_format | dspace |
| spelling | upm.eprints-164392015-09-23T07:56:40Z http://psasir.upm.edu.my/id/eprint/16439/ Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis Kesar Singh, Jasbeer Kaur Radu, Son Mohamad Ghazali, Farinazleen Cheah, Yoke Kqueen Successful DNA amplification is vital for the detection of specific DNA targets in feeds, and this in return depends on the ability of DNA extraction methods to produce good quality DNA. In this study, seven methods were compared for DNA extraction from feeds using quantitative polymerase chain reaction (PCR) of single copy maize (Zea mays) endogenous hmg (high mobility group) gene. Relative levels of hmg were used to evaluate the DNA quality. Spectrophotometer determination of DNA was also carried out to assess DNA yield and DNA purity, while electrophoretic analysis of genomic DNA extracts was carried out to investigate DNA integrity. The findings illustrate that the DNA extraction methods have a significant effect on DNA quality. Statistically, the Epicentre method extracted the highest DNA yield while the Wizard method had the lowest DNA yield with high DNA purity and integrity. However, the Wizard method recovered the most amplifiable DNA per reaction, indicating that template quality and integrity had greater influence over hmg amplification than DNA yield. Faculty of Food Science and Technology, Universiti Putra Malaysia 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16439/1/16439.pdf Kesar Singh, Jasbeer Kaur and Radu, Son and Mohamad Ghazali, Farinazleen and Cheah, Yoke Kqueen (2009) Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis. International Food Research Journal, 16 (3). pp. 329-341. ISSN 1985-4668; ESSN: 2231-7546 http://www.ifrj.upm.edu.my/16%20%283%29%202009/6.pdf |
| spellingShingle | Kesar Singh, Jasbeer Kaur Radu, Son Mohamad Ghazali, Farinazleen Cheah, Yoke Kqueen Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title | Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title_full | Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title_fullStr | Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title_full_unstemmed | Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title_short | Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis |
| title_sort | real time pcr evaluation of seven dna extraction methods for the purpose of gmo analysis |
| url | http://psasir.upm.edu.my/id/eprint/16439/1/16439.pdf |
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