Affiliation: | 1.Department of Biomedical Engineering, Faculty of Life Sciences,University of Applied Sciences Albstadt-Sigmaringen,Sigmaringen,Germany;2.State Institute of Chemical and Veterinarian Analysis of Food Sigmaringen (CVUA Sigmaringen),Sigmaringen,Germany |
Abstract: | Performance of DNA-based methods used for allergen detection does not only depend on the real-time PCR system, but also on the DNA isolation method used. Comparison of different isolation methods showed that yield and purity of isolated DNA strongly depends on the isolation method deployed as well as on the composition of the food matrix (matrix effect). Detection of trace amounts of allergens requires efficient methods for DNA isolation which remove inhibitors and yield pure DNA. This is particularly important for quantitative analysis. Due to matrix effects, methods, which have the ability to circumvent these effects, are of great interest. Therefore, a sequence-specific method for isolation of target DNA based on magnetic particles (MCH) has been established and compared to other methods for isolation of total DNA. |