DNA and RNA obtained from Bouin's fixed tissues

Author(s): Bonin S, Petrera F, Rosai J, Stanta G

Publication: J Clin Pathol, 2005, Vol. 58, Page 313

PubMed ID: 15735167 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   To determine if DNA extracted from BFPE breast cancer biopsies is sufficient in abundance and quality for PCR analysis.

   Summary of Findings:

   Successful PCR amplification of three target genes (epidermal growth factor receptor (EGFR), p16, transthyretin (TTR)) was improved (with shorter amplicons) or contingent (with longer amplicons) upon an additional restoration step. In samples that underwent restoration, PCR amplification success was inversely proportional to the targeted product size. PCR amplification failed in samples that did not undergo restoration for larger target product sizes.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   PCR Specific	Length of gene fragment	90 bp 204 bp 291 bp 333 bp

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2. Study Purpose

   To determine if RNA extracted from BFPE breast cancer biopsies is sufficient in abundance and quality for RT-PCR analysis, and whether amplicon size impacts PCR success.

   Summary of Findings:

   RT-PCR success for beta actin, EGFR, and GAPDH mRNA expression was inversely proportional to amplicon size, with a 100% success rate for 65-75 bp generated amplicons which decreased to 80% for 103 bp amplicons. No product was observed for targeted products of 170 bp or greater.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   RT-PCR Specific	Length of gene fragment	65 bp 75 bp 77 bp 95 bp 100 bp 103 bp 120 bp 170 bp 200 bp

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