An integrated microsatellite length analysis using an automated fluorescent DNA sequencer

Yasushi Toh, Eiji Oki, Shinya Oda, Masaaki Tomoda, Shinichi Tomisaki, Yuji Ichiyoshi, Shinji Ohno, Keizo Sugimachi

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Analyzing microsatellite instability (MI) in malignant tumors is thought in be useful for screening cancer patients to identify those patients with a higher risk of developing second malignant tumors. In this paper, we report a new, accurate, and efficient method of detecting MI using an automated fluorescent DNA sequencer and a computer that automatically calculates the size, height, and area of each fluorescent product, making it possible to assess MI more accurately and more rapidly. The primers for amplification of each microsatellite locus are labeled by two different fluorescent dyes, rox (red) and fam (blue). The rox-labeled primer was used for the tumor, whereas the fam-labeled primer was used for the corresponding normal tissue. Two amplified products from both the tumor and the normal tissue were co-loaded into a single lane of the sequencing gel and were analyzed. MI could be detected based on the presence of different waving patterns. Furthermore, several loci could also be analyzed simultaneously for MI in a single lane. Using this method, we examined the frequency of MI in gastric cancer. The results showed that 5 of 22 (22.7%) gastric cancers were MI-positive, which corresponds to the findings of previous reports that used the radioisotopic method. The improved method may open up the possibility of performing routine examination of MI in many cancer patients and offers hope for the potential clinical application of MI analysis as a follow-up evaluation of cancer patients.

Original languageEnglish
Pages (from-to)2688-2691
Number of pages4
JournalCancer Research
Volume56
Issue number12
Publication statusPublished - Jun 15 1996

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Microsatellite Instability
Microsatellite Repeats
DNA
Neoplasms
Stomach Neoplasms
Fluorescent Dyes
Early Detection of Cancer
Gels

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Toh, Y., Oki, E., Oda, S., Tomoda, M., Tomisaki, S., Ichiyoshi, Y., ... Sugimachi, K. (1996). An integrated microsatellite length analysis using an automated fluorescent DNA sequencer. Cancer Research, 56(12), 2688-2691.

An integrated microsatellite length analysis using an automated fluorescent DNA sequencer. / Toh, Yasushi; Oki, Eiji; Oda, Shinya; Tomoda, Masaaki; Tomisaki, Shinichi; Ichiyoshi, Yuji; Ohno, Shinji; Sugimachi, Keizo.

In: Cancer Research, Vol. 56, No. 12, 15.06.1996, p. 2688-2691.

Research output: Contribution to journalArticle

Toh, Y, Oki, E, Oda, S, Tomoda, M, Tomisaki, S, Ichiyoshi, Y, Ohno, S & Sugimachi, K 1996, 'An integrated microsatellite length analysis using an automated fluorescent DNA sequencer', Cancer Research, vol. 56, no. 12, pp. 2688-2691.
Toh Y, Oki E, Oda S, Tomoda M, Tomisaki S, Ichiyoshi Y et al. An integrated microsatellite length analysis using an automated fluorescent DNA sequencer. Cancer Research. 1996 Jun 15;56(12):2688-2691.
Toh, Yasushi ; Oki, Eiji ; Oda, Shinya ; Tomoda, Masaaki ; Tomisaki, Shinichi ; Ichiyoshi, Yuji ; Ohno, Shinji ; Sugimachi, Keizo. / An integrated microsatellite length analysis using an automated fluorescent DNA sequencer. In: Cancer Research. 1996 ; Vol. 56, No. 12. pp. 2688-2691.
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