TY - JOUR
T1 - Simplified procedures for applying the polymerase chain reaction to routinely fixed paraffin wax sections
AU - Coates, P. J.
AU - d'Ardenne, A. J.
AU - Khan, G.
AU - Kangro, H. O.
AU - Slavin, G.
PY - 1991
Y1 - 1991
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0026058238&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026058238&partnerID=8YFLogxK
U2 - 10.1136/jcp.44.2.115
DO - 10.1136/jcp.44.2.115
M3 - Article
C2 - 1650795
AN - SCOPUS:0026058238
SN - 0021-9746
VL - 44
SP - 115
EP - 118
JO - Journal of Clinical Pathology
JF - Journal of Clinical Pathology
IS - 2
ER -