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

• Published in: Journal of clinical pathology Vol. 44; no. 2; pp. 115 - 118
• Main Authors: Coates, P J, d'Ardenne, A J, Khan, G, Kangro, H O, Slavin, G
• Format: Journal Article
• Language: English
• Published: London BMJ Publishing Group Ltd and Association of Clinical Pathologists 01-02-1991; BMJ; BMJ Publishing Group LTD
• Subjects: Analytical, structural and metabolic biochemistry; Base Sequence; Biological and medical sciences; Culture Techniques; DNA, Viral - analysis; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Herpesvirus 4, Human - isolation & purification; Histological Techniques; Humans; Molecular Sequence Data; Nasopharyngeal Neoplasms - microbiology; Nucleic acids; Oligonucleotides; Paraffin; Polymerase Chain Reaction - methods; Fixation; DNA; Extraction; Method; Paraffin; Biological material; Quantitative analysis
• ISSN: 0021-9746; 1472-4146
• DOI: 10.1136/jcp.44.2.115

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.