Analysis of microsatellite instability by microfluidic-based electrophoresis.

ABSTRACT

Microsatellites are short repetitive sequences of two, three, or four bases, prone to base mispairing. Microsatellite instability (MSI) occurs frequently in various types of cancer due to a defective DNA mismatch repair system. Therefore, MSI analysis is an important tool in clinical research and molecular diagnostics. Mostly, polyacrylamide gel electrophoresis or capillary electrophoresis of labeled microsatellite sequences is used for the detection of MSI. Here we present a microfluidic-based electrophoresis technology for MSI analyses. Defined loci of microsatellites were PCR amplified and amplicons were analyzed by microfluidic-based electrophoresis. The electropherogram profiles of tumor and non-tumor derived DNA clearly revealed an individual pattern identifying differences in tumor-associated microsatellites. Detection of MSI by microfluidics turned out to be a simple and efficient procedure but less laborious than conventional approaches. Thus, the chip-based microfluidic electrophoresis is a simple, reliable, and robust technology for MSI detection, which allows label-free analyses of microsatellite amplicons within 30 min.