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PMID: 1650795 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Simplified procedures for applying the polymerase chain reaction to routinely fixed paraffin wax sections.

Journal of clinical pathology ·Vol. 44 ·No. 2 ·1991-02-00 ·Pages 115-8

Coates PJ, d'Ardenne AJ, Khan G, Kangro HO, Slavin G

Abstract

The polymerase chain reaction was applied to the analysis of DNA contained in archival paraffin wax embedded material. DNA suitable for the reaction was obtained from these tissues by simple extraction methods, without previous dewaxing of tissue sections. When compared with unfixed material, the reaction efficiency was compromised, so that an increased number of amplification cycles were required to produce equivalent amounts of amplified product. This in turn led to an increase in amplification artefacts, which can be minimised by a simple modification of the standard reaction. Amplification of relatively large DNA fragments was not always successful, and it seems prudent to bear this in mind when designing oligonucleotide primers which are to be used for the amplification of archival material. The efficiency of the procedure can be improved by dividing the amplification cycles into two parts: this reduces the amount of reagent needed, is relatively simple and inexpensive, and can be performed in one working day.

MeSH Terms
Base Sequence Culture Techniques DNA, Viral/analysis Electrophoresis, Agar Gel Herpesvirus 4, Human/isolation & purification Histological Techniques Humans Molecular Sequence Data Nasopharyngeal Neoplasms/microbiology Oligonucleotides Paraffin Polymerase Chain Reaction/methods
Chemicals
DNA, Viral Oligonucleotides Paraffin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Coates P J
Department of Histopathology, St Bartholomew's Hospital, West Smithfield, London.
d'Ardenne A J
Khan G
Kangro H O
Slavin G
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Article Info
Journal
Journal of clinical pathology
Abbr.
J Clin Pathol
ISSN
0021-9746
Published
1991-02-00
Pages
115-8
Language
English
Region
England
NLM ID
0376601
PMCID
PMC496971
Subset
IM
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