Article
Lab Invest 2002, 82:1707–1714
Construction of cDNA Libraries from Microdissected Benign and Malignant Thyroid Tissue
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Klaus Kaserer1, Vladimir Knezevic3, Bettina Pichlhöfer1, Christian Scheuba2, Christian Passler2, Jennifer Worth4, Bruno Niederle2 and David Krizman4
- 1Department of Clinical Pathology, University of Vienna Medical School, Vienna, Austria
- 2Department of Surgery, University of Vienna Medical School, Vienna, Austria
- 320/20 Gene Systems, Inc., Rockville, Maryland
- 4Office of the Director, The Cancer Genome Anatomy Project, National Cancer Institute, National Institutes of Health, Bethesda, Maryland
Correspondence: Dr. K. Kaserer, Department of Clinical Pathology, University of Vienna Medical School, Währinger Gürtel 18-20, A-1090 Vienna, Austria. E-mail: k.kaserer@akh-wien.ac.at
Received 11 September 2002.
Abstract
cDNA libraries were constructed from thyroid epithelial cells gained by laser capture microdissection for gene expression analysis of the progression of thyroid cancer. Six histologically diverse thyroid tissue specimens were used. A mean of 93 ng of total RNA was gained per tissue sample from a mean estimated number of 25,000 microdissected cells per sample. Analysis of randomly selected clones from six libraries showed an average insert size of 600 (range, 300–1500) bp. Preliminary sequencing of clones selected from the six libraries indicates a range of 46% to 62% known genes per library, 4% to 25% anonymous expressed sequence tags per library, and 15% to 43% novel expressed sequence tags per library. Thyroglobulin was found in normal thyroid epithelium and follicular thyroid adenoma, whereas calcitonin precursor transcripts were found in medullary thyroid carcinoma. We demonstrate production of high-quality cDNA libraries of microdissected tissue of the thyroid, which should prove useful for gene expression analysis of human thyroid tumors.
