Real-time PCR evaluation of seven DNA extraction methods for the purpose of GMO analysis
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
Official URL: http://www.ifrj.upm.edu.my/volume-16-2009.html
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.
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