Construction of a representative cDNA library from prostatic intraepithelial neoplasia

David B. Krizman, Rodrigo F. Chuaqui, Paul S. Meltzer, Jeffrey M. Trent, Paul H. Duray, W. Marston Linehan, Lance A. Liotta, Michael R. Emmert-Buck

Research output: Contribution to journalArticle

Abstract

We report the construction of a plasmid-based cDNA library made from microdissected cells derived from prostatic intraepithelial neoplasia. Total RNA was extracted and converted to blunt-ended, double-stranded cDNA by oligo(dT)-mediated reverse transcription followed by linker addition. A linker-specific primer with UDG-compatible ends was used to amplify the cDNA and the resulting PCR product was subcloned. A total of 154 clones were sequenced and results indicated that 81.5% of the clones derived from either known genes, anonymous expressed sequence tags, or novel transcripts with very little redundancy of screened clones. These results demonstrate the feasibility of constructing complex representative cDNA libraries from specific microdissected cell populations that represent microscopic precursor stages of cancer progression. This method should facilitate identification of transcripts specifically expressed in cells of a distinct histological origin and tumorigenic stage.

Original languageEnglish (US)
Pages (from-to)5380-5383
Number of pages4
JournalCancer Research
Volume56
Issue number23
StatePublished - Dec 1 1996
Externally publishedYes

Fingerprint

Prostatic Intraepithelial Neoplasia
Gene Library
Clone Cells
Complementary DNA
Expressed Sequence Tags
Reverse Transcription
Plasmids
RNA
Polymerase Chain Reaction
Population
Genes
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Krizman, D. B., Chuaqui, R. F., Meltzer, P. S., Trent, J. M., Duray, P. H., Linehan, W. M., ... Emmert-Buck, M. R. (1996). Construction of a representative cDNA library from prostatic intraepithelial neoplasia. Cancer Research, 56(23), 5380-5383.

Construction of a representative cDNA library from prostatic intraepithelial neoplasia. / Krizman, David B.; Chuaqui, Rodrigo F.; Meltzer, Paul S.; Trent, Jeffrey M.; Duray, Paul H.; Linehan, W. Marston; Liotta, Lance A.; Emmert-Buck, Michael R.

In: Cancer Research, Vol. 56, No. 23, 01.12.1996, p. 5380-5383.

Research output: Contribution to journalArticle

Krizman, DB, Chuaqui, RF, Meltzer, PS, Trent, JM, Duray, PH, Linehan, WM, Liotta, LA & Emmert-Buck, MR 1996, 'Construction of a representative cDNA library from prostatic intraepithelial neoplasia', Cancer Research, vol. 56, no. 23, pp. 5380-5383.
Krizman DB, Chuaqui RF, Meltzer PS, Trent JM, Duray PH, Linehan WM et al. Construction of a representative cDNA library from prostatic intraepithelial neoplasia. Cancer Research. 1996 Dec 1;56(23):5380-5383.
Krizman, David B. ; Chuaqui, Rodrigo F. ; Meltzer, Paul S. ; Trent, Jeffrey M. ; Duray, Paul H. ; Linehan, W. Marston ; Liotta, Lance A. ; Emmert-Buck, Michael R. / Construction of a representative cDNA library from prostatic intraepithelial neoplasia. In: Cancer Research. 1996 ; Vol. 56, No. 23. pp. 5380-5383.
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