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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Frayling IM (1999) Microsatellite instability. Gut 45:1–4
Grady WM (2004) Genomic instability and colon cancer. Cancer Metastasis Rev 23:11–27
Muc R, Naidoo R (2002) Microsatellite instability in diagnostic pathology. Curr Diagn Pathol 8:318–327
Berg KD et al (2000) Detection of microsatellite instability by fluorescence multiplex polymerase chain reaction. J Mol Diagn 2:20–28
Ugaz VM et al (2004) Microfabricated electrophoresis systems for DNA sequencing and genotyping applications: current technology and future directions. Philos Transact A Math Phys Eng Sci 362:1105–1129
Banerjea A et al (2003) Colorectal cancers with mononucleotide microsatellite instability can be identified using microfabricated chip technology. Anal Biochem 322:130–133
Odenthal M et al (2009) Analysis of microsatellite instability in colorectal carcinoma by microfluidic-based chip electrophoresis. J Clin Pathol 62:850–852
Dietmaier W et al (1997) Diagnostic microsatellite instability: definition and correlation with mismatch repair protein expression. Cancer Res 57:4749–4756
Agilent DNA 1000 kit: quick start guide. Manual of the Agilent DNA 1000 kit; Agilent Biotechnologies, Waldbronn, Germany
Acknowledgments
We thank Rüdiger Salowsky (Agilent Technologies, Waldbronn, Germany) for his technical advice and support in the evaluation of microfluidic-based electrophoresis.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Elfimova, N., Amer, W., Odenthal, M. (2013). Analysis of Microsatellite Instability by Microfluidic-Based Electrophoresis. In: Phillips, T., Kalish, H. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 919. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-029-8_25
Download citation
DOI: https://doi.org/10.1007/978-1-62703-029-8_25
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-028-1
Online ISBN: 978-1-62703-029-8
eBook Packages: Springer Protocols