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Mutational Analysis of the Androgen Receptor Using Laser Capture Microdissection and Direct Sequencing

  • Sheila Greene
  • Patricia Stockton
  • Olga G. Kozyreva
  • Kris E. Gaston
  • Gordon P. Flake
  • James L. Mohler
Part of the Methods in Molecular Biology™ book series (MIMB, volume 218)

Abstract

Molecular analysis of prostate cancer specimens is complicated by tumor heterogeneity and admixture of malignant cells with benign epithelium, stroma and inflammatory cells. Laser Capture Microdissection allows procurement of selected groups of cells from specific areas of tissue sections. We have used Laser Capture Microdissection to obtain pure samples of malignant prostate epithelial cells from frozen radical prostatectomy specimens. Methods for DNA extraction, androgen receptor amplification and exon sequencing have been optimized. These methods should be adaptable for molecular analysis of DNA, RNA and protein from other complex tissues.

Key Words

Laser capture microdissection androgen receptor mutations prostate cancer polymerase chain reaction direct sequencing 

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Copyright information

© Humana Press Inc., Totowa, NJ 2003

Authors and Affiliations

  • Sheila Greene
    • 1
  • Patricia Stockton
    • 2
  • Olga G. Kozyreva
    • 1
  • Kris E. Gaston
    • 1
    • 3
  • Gordon P. Flake
    • 2
  • James L. Mohler
    • 1
    • 3
  1. 1.Division of Urology, Department of SurgeryUniversity of North Carolina at Chapel HillChapel Hill
  2. 2.Laboratory of Experimental PathologyNational Institute of Environmental Health SciencesResearch Triangle Park
  3. 3.UNC-Lineberger Comprehensive Cancer CenterUniversity of North Carolina at Chapel HillChapel Hill

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