Original Articles

Annals of Surgical Oncology

, Volume 7, Issue 4, pp 276-280

Diagnostic Primer Sets for Microsatellite Instability Optimized for a Minimal Amount of Damaged DNA From Colorectal Tissue Samples

  • Naoyuki UmetaniAffiliated withDepartment of Surgical Oncology, School of Medicine, University of TokyoDepartment of Surgical Oncology, School of Medicine, University of Tokyo Email author 
  • , Shin SasakiAffiliated withDepartment of Surgical Oncology, School of Medicine, University of Tokyo
  • , Toshiaki WatanabeAffiliated withDepartment of Surgical Oncology, School of Medicine, University of Tokyo
  • , Hironori IshigamiAffiliated withDepartment of Surgical Oncology, School of Medicine, University of Tokyo
  • , Eiji UedaAffiliated withDepartment of Surgical Oncology, School of Medicine, University of Tokyo
  • , Hirokazu NagawaAffiliated withDepartment of Surgical Oncology, School of Medicine, University of Tokyo

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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.

Key Words:

Colorectal carcinoma—Microdissection—PCR—Microsatellite instability.