The influence of age of template DNA derived from archival tissue on the outcome of the polymerase chain reaction.
The analysis of DNA from archival tumour tissue for molecular alterations has been facilitated by the use of the polymerase chain reaction (PCR). Degradation of tissue prior to fixation and the nature of the fixative used influence successful amplification from archival tissue. Age of the archival tissue may also be a factor. To determine if this was so, DNA was extracted from 30 archival specimens of spleen spanning a 15 year period. Polymerase chain reaction was performed on all specimens using primers for exon 2 (307 bp) and exon 9 (1278 bp) of the hypoxanthine phosphoribosyl transferase (HPRT) gene. It was not possible to show that the age of archival tissue had an influence on the capacity to amplify exon 2 of the HPRT gene. It was not possible to amplify exon 9 of the HPRT gene from archival tissue.
Hewett
P J
PJ
Department of Surgery, Flinders Medical Centre, Bedford Park, South Australia.
Firgaira
F
F
Morley
A
A
D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D005091:Exons / Q000235:genetics; D005404:Fixatives; D005557:Formaldehyde; D005784:Gene Amplification; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D013154:Spleen / Q000737:chemistry; D013997:Time Factors; D016707:Tissue Fixation
D004273:DNA, Neoplasm; D005404:Fixatives; D005557:Formaldehyde; D007041:Hypoxanthine Phosphoribosyltransferase