Abstract
Background: The diagnosis of microsatellite instability from a minimal amount of highly damaged DNA, extracted from formalin-fixed, paraffin-embedded tissue by the microdissection method, is difficult. Therefore, optimized primer sets were newly designed for substitution of documented ones.
Methods: DNA was extracted from 15 archival colorectal carcinomas and used as templates for polymerase chain reaction. Nine standard microsatellite markers (BAT-25, BAT-26, BAT-40, D18S69, D2S123, D5S346, D10S197, D17S250, and D18S58) were selected for diagnosis of microsatellite instability in colorectal carcinomas. All polymerase chain reaction conditions for primer sets were unified to save experimental time.
Results: The primer sets for the latter five markers documented in the literature were redesigned because of poor efficiency for damaged DNA. As a result, the number of DNA samples, sufficiently amplified at all markers, improved from 0% to 93%.
Conclusions: Diagnostic primer sets for microsatellite instability, optimized for a minimal amount of damaged DNA from colorectal tissue samples, were established.
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Umetani, N., Sasaki, S., Watanabe, T. et al. Diagnostic Primer Sets for Microsatellite Instability Optimized for a Minimal Amount of Damaged DNA From Colorectal Tissue Samples. Ann Surg Oncol 7, 276–280 (2000). https://doi.org/10.1007/s10434-000-0276-6
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DOI: https://doi.org/10.1007/s10434-000-0276-6