Automated Genotyping With Fluorescence Correlation Spectroscopy

Both the MF10S microplate reader [developed jointly by Olympus Optical Co., LTD (Tokyo, Japan) and Evotec

Technologies GmbH] and the Analyzer AA2.0 (Evotec Technologies GmbH) are bench-top instruments for automated FCS analysis and perform automated adjustment, FCS measurements, autocorrelation, data fitting, and SNP genotype assignment. Reaction mixes are excited at 543 and 633 nm, and the quantities of allele-specific fluorescent amplification products (% amplimer) at each wavelength are determined. Genotypes are assigned by calculation of a normalized fluorescence signal (NFS), i.e., the difference of the quantities of fluorescent amplification products normalized by the sum of all amplimer-derived signal [(543 - 633)/(543 + 633)]. The software employs user-defined or default limit values for multiple fluorescence parameters and the quantities of fluorescent amplification products from negative and positive control reactions, thus providing multiple intrinsic quality controls for successful amplification and single molecule detection in each reaction. Additionally, user-defined limits for discrimination of the three possible genotypes allow for automated SNP genotype assignment.

Was this article helpful?

0 0
Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

Get My Free Ebook

Post a comment