A method to isolate DNA from small archival tissue samples for p53 gene analysis

Abstract

The tumor suppressor gene p53 is involved in predisposition to a variety of human cancers, including those from Li-Fraumeni cancer family syndrome patients. Studies of inheritance of p53 germline mutations require confirmation of the mutation in the tumors from family members. These studies, as well as other retrospective studies of tumor specific mutations, are often hampered by a lack of available fresh or frozen tumor tissue samples for DNA extraction to confirm the suspected p53 mutation. Here we describe a simple technique for DNA isolation that permits mutational analysis of p53 from minimal amounts of paraffin-embedded archival tissue samples.

Detection of DNA by scraping the formalin-fixed paraffin-embedded tissue

“…Keeping in mind this problem, other scientists have also adopted different techniques to overcome this for DNA isolation to permit mutational analysis of p53 from minimal amounts of paraffin-embedded archival tissue samples. The authors followed Chen and Clejan [12], Schubert et al [7], and Koyama et al [8], who introduced very simple methods for DNA detection from formalin-fixed paraffin-embedded tissues but these techniques are still cumbersome to perform in an area with limited facilities and for the research students in universities of Pakistan. The authors modified the procedures to make them easier for local use.…”

“…The purification of DNA from paraffin-embedded sections of human squamous cell carcinoma has been successfully used to detect the prevalence and *Corresponding author: 26-MOF, GOR-3, Shadman, Lahore, 54000, Pakistan. email: drmhbukhari@yahoo.com localization of HPV using a modified in situ polymerase chain reaction (PCR) and in situ hybridization AT tailing (ISH-AT) [7,8]. The Gentra system is an important method for genomic DNA purification and is known as the Puregene DNA purification system.…”

A germline missense mutation R337C in exon 10 of the human p53 gene

“…DNA was isolated from paraffin-embedded tissue as previously described (Schubert et al, 1993). The DNA was amplified by polymerase chain reaction (PCR) using previously described primers (Mashiyama et al, 1991).…”

Evidence for a Novel Osteosarcoma Tumor-Suppressor Gene in the Chromosome 18 Region Genetically Linked with Paget Disease of Bone

“…The specimens from the Paget disease patients were obtained as formalin-fixed samples. In these cases, DNA was isolated as described elsewhere for paraffin-embedded tumor samples (Schubert et al 1993).…”