RT-PCR

Variations in the human genome are analysed on an Applied Biosystems real time PCR instrument. DNA from buccal swaps or blood samples is extracted and specifically designed primers for the base sequence of interest bind to the corresponding DNA region. Flurophores added to the primers are used to monitor the binding process.

LCMS-Q-TOF

A Bruker Daltonics microTOF-Q coupled to a Dionex Ultimate 3000 micro-HPLC-system provides mass spectra with high mass accuracy and MS/MS-capabilities for metabolite identification and high sensitivity and dynamic range for metabolite quantification. This technique enables us to assess a wide range of metabolites including unknown compounds which may be affected by the genetic background. This is in contrast to classical LC-MS, which only allows identification of specific metabolites which need to be identified prior to method development.

PCA

The enormous amount of data generated by LC-MS/MS is processed by metabolic profiler software to allow data clustering and post-run evaluation by principle component analysis. The software is run in close collaboration with Bruker. This enables us to identify metabolic variations by combining characteristic modifications and excluding extrinsic influences like diet, physical activity, or any other environmental condition interfering with the interventional parameters.