Application of polymerase chain reaction-restriction fragment length polymorphism analysis and lab-on-a-chip capillary electrophoresis for the specific identification of game and domestic meats
2009
Fajardo, Violeta | González, Isabel | Dooley, John | Garret, Steve | Brown, H. M. (Helen M) | García, Teresa | Martín, Rosario
BACKGROUND: The objective of this study was to adapt and improve previously published polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods aimed at the identification of game and domestic meats, by replacing the gel electrophoretic steps for DNA fragment analysis by a chip-based capillary electrophoresis system.RESULTS: PCR amplification of a mitochondrial 12S rRNA gene fragment and subsequent digestion of the amplicons with either MseI or a combination of MboII, BslI, and ApoI endonucleases generated characteristic PCR-RFLP profiles that allowed discrimination among ten relevant game and domestic meat species. The Agilent 2100 Bioanalyzer utilises capillary electrophoresis on a microchip device that is capable of rapidly sizing DNA fragments, offering a valuable recent development for the analysis of complex DNA banding patterns.CONCLUSION: Results obtained in this work indicated that banding resolution on the system was sensitive, with detection of some small DNA fragments that were not observed with the published conventional PCR-RFLP gel-based method. Therefore, the new faster and easy handling procedure provides an additional powerful tool that can be employed for meat speciation.
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