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Related Experiment Videos

Whole-genome allelotyping using laser microdissected tissue.

Colleen M Feltmate1, Samuel C Mok

  • 1Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 21, 2005
PubMed
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Laser-based microdissection enables tumor cell isolation for DNA analysis. This technique, combined with whole-genomic amplification, allows for high-throughput genome-wide allelotyping and loss of heterozygosity assessment.

Area of Science:

  • Molecular Biology
  • Genetics
  • Oncology

Background:

  • Laser-based microdissection (LBM) is a novel technology.
  • LBM can procure homogenous tumor cells from tissue sections.
  • Tumor cells are often limited in quantity.

Purpose of the Study:

  • To evaluate LBM for tumor cell isolation.
  • To assess the feasibility of whole-genomic amplification (WGA) on LBM-procured cells.
  • To perform genome-wide allelotyping on amplified DNA.

Main Methods:

  • Laser-based microdissection of tumor cells from paraffin-embedded and frozen sections.
  • Whole-genomic amplification of DNA from microdissected cells.
  • High-throughput genome-wide allelotyping using fluorescent-labeled microsatellite markers.

Related Experiment Videos

  • Analysis of loss of heterozygosity using Genescan and Genotyper software.

    • Main Results:

    • Sufficient DNA was generated from small numbers of tumor cells using LBM and WGA.
    • Genome-wide allelotyping was successfully performed on amplified DNA.
    • Loss of heterozygosity patterns were assessed.

    Conclusions:

    • LBM combined with WGA is a viable method for obtaining sufficient DNA from limited tumor samples.
    • This approach facilitates high-throughput genome-wide allelotyping for cancer research.
    • The methodology allows for the assessment of loss of heterozygosity in tumor cells.