Microwave-based DNA extraction from paraffin-embedded tissue for PCR amplification.

Author(s): Banerjee SK, Makdisi WF, Weston AP, Mitchell SM, Campbell DR

Publication: Biotechniques, 1995, Vol. 18, Page 768-70, 772-3

PubMed ID: 7619474 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of deparaffinization, proteinase k treatment and DNA extraction method on the amplification of ki-ras from formaldehyde-fixed paraffin-embedded tissue sections.

   Summary of Findings:

   When DNA was extracted by microwave irradiation followed by proteinase k digestion, amplification of ki-ras was as successful as when DNA was isolated by xylene deparaffinization, proteinase k digestion and phenol-chloroform extraction. Phenol-chloroform extraction after proteinase k digestion did not improve the amplification of ki-ras compared to simple boiling, but the proteinase k digestion step was necessary to maintain yield. The authors report that proteinase k digestion was effective when performed for 3 h at 55 degrees C instead of overnight at 42 degrees C.

   Biospecimens

   • Tissue - Pancreas

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Deparaffinization	Microwave Xylene
   Analyte Extraction and Purification	Protein digestion	None Proteinase K Overnight at 42 degrees C 3 h at 55 degrees C
   Analyte Extraction and Purification	Analyte isolation method	Boiling Phenol-chloroform extraction

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