Abstract
Microsatellite analysis is a frequently used method for detection of chromosomal deletions by loss of heterozygosity studies and for detection of microsatellite instability. For reliable microsatellite analyses, a tumor cell content of at least 80% is required. Therefore, laser microdissection is an important prerequisite for those studies, allowing the contamination-free isolation of morphologically defined pure tumor cell populations. The combination of exact microdissection and subsequent whole-genome amplification by improved primer extension preamplification polymerase chain reaction (I-PEP-PCR) facilitates the analysis of multiple microsatellite loci in small tumor samples. This is especially important for the investigation of malignant tumors with low tumor cellularity. This chapter describes in detail the use of whole genome amplification by I-PEP PCR and microsatellite analysis in laser microdissected specimens of colon and breast cancer.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Dietmaie, W., Wallinge, S., Bocker, T., Kullmann, F., Fishel, R., and Rüschoff, J. (1997) Diagnostic microsatellite instability: Definition and correlation with mismatch repair protein expression. Cancer Res. 57,4749–4756.
Boland, C. R., Thibodeau, S. N., Hamilton, S. R., et al. (1998) A National Cancer Institute Workshop on Microsatellite Instability for cancer detection and familial predisposition: Development of international criteria for the determination of microsatellite instability in colorectal cancer. Cancer Res. 58, 5248–5257.
Ahrendt, S. A., Decker, P. A., Doffek, K., et al. (2000) Microsatellite instability at selected tetranucleotide repeats is associated with p53 mutations in non-small cell lung cancer. Cancer Res. 60, 2488–2491.
Dietmaier, W., Hartmann, A., Wallinger, S., et al. (1999) Multiple mutation analyses in single tumor cells with improved whole genome amplification. Am. J.Pathol. 154, 883–895.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Wild, P.J., Stoehr, R., Knuechel, R., Hartmann, A., Dietmaier, W. (2005). Laser Microdissection for Microsatellite Analysis in Colon and Breast Cancer. In: Murray, G.I., Curran, S. (eds) Laser Capture Microdissection. Methods in Molecular Biology™, vol 293. Humana Press. https://doi.org/10.1385/1-59259-853-6:093
Download citation
DOI: https://doi.org/10.1385/1-59259-853-6:093
Publisher Name: Humana Press
Print ISBN: 978-1-58829-260-5
Online ISBN: 978-1-59259-853-3
eBook Packages: Springer Protocols