Simultaneous assessment of loss of heterozygosity at multiple microsatellite loci using semi-automated fluorescence-based detection: Subregional mapping of chromosome 4 in cervical carcinoma Academic Article uri icon

Overview

MeSH Major

  • Chromosome Deletion
  • Chromosomes, Human, Pair 4
  • DNA, Satellite
  • Heterozygote
  • Microsatellite Repeats
  • Uterine Cervical Neoplasms

abstract

  • Detection of loss of heterozygosity (LOH) by comparison of normal and tumor genotypes using PCR-based microsatellite loci provides considerable advantages over traditional Southern blotting-based approaches. However, current methodologies are limited by several factors, including the numbers of loci that can be evaluated for LOH in a single experiment, the discrimination of true alleles versus "stutter bands," and the use of radionucleotides in detecting PCR products. Here we describe methods for high throughput simultaneous assessment of LOH at multiple loci in human tumors; these methods rely on the detection of amplified microsatellite loci by fluorescence-based DNA sequencing technology. Data generated by this approach are processed by several computer software programs that enable the automated linear quantitation and calculation of allelic ratios, allowing rapid ascertainment of LOH. As a test of this approach, genotypes at a series of loci on chromosome 4 were determined for 58 carcinomas of the uterine cervix. The results underscore the efficacy, sensitivity, and remarkable reproducibility of this approach to LOH detection and provide subchromosomal localization of two regions of chromosome 4 commonly altered in cervical tumors.

publication date

  • June 25, 1996

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC39090

PubMed ID

  • 8692882

Additional Document Info

start page

  • 6704

end page

  • 9

volume

  • 93

number

  • 13