Analysis of DNA extracted from formalin-fixed, paraffin-embedded tissues by enzymatic amplification and hybridization with sequence-specific oligonucleotides

doi:10.1016/0006-291X(87)91472-0

Biochemical and Biophysical Research Communications

Volume 142, Issue 3, 13 February 1987, Pages 710-716

https://doi.org/10.1016/0006-291X(87)91472-0 Get rights and content

Abstract

The “polymerase chain reaction” (PCR) procedure for amplifying specific gene sequences has recently been combined with sequence-specific oligonucleotide (SSO) probe hybridization to develop a highly sensitive, rapid, and simple method for analyzing allelic variations in genomic DNA (1). In the present study we have used $PCR SSO$ to analyze partially purified DNA extracted from formalin-fixed, paraffin-embedded tissue specimens. We report that this DNA, including samples that were partially degraded, proved to be suitable for analysis by the $PCR SSO$ procedure.

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The first approach is DNA analysis. However, egg samples are fixed with formalin to preserve their morphological characteristics and this results in DNA fragmentation and protein cross-linking, which makes DNA extraction difficult or impossible (e.g., Goelz et al., 1985; Impraim et al., 1987). The second approach is to use a mixture distribution of the eggs of chub mackerel and spotted mackerel which contains the temporal changes in egg diameters of two species.
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Subclinical infection can be detected by colposcopy, while latent infection can only be diagnosed using molecular biology techniques, since such infection is not accompanied by clinical symptoms [8]. The prevalence of HPV in anal tissue without neoplasia has ranged from 0% [9] to 16.7% [10], while the reported prevalence of this same virus associated with anal carcinoma has been as high as 84.6% [11]. The presence of HPV in the uterine cervix has been associated with the presence of this virus in the anus.
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This study was approved by the Institutional Ethics Committee, and all patients signed an informed consent document. Genomic DNA was extracted from formalin-fixed and paraffin-embedded tissues by the proteinase K-phenol-chloroform extraction method.19 All human DNA samples were examined using the primers from a cellular gene hypoxanthine-guanine phosphoribosyl transferase (HGPRT).20
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Analysis of DNA extracted from formalin-fixed, paraffin-embedded tissues by enzymatic amplification and hybridization with sequence-specific oligonucleotides

Abstract

Biochem Biophys Res Commun

Nature

Am J Human Genet

Cancer Res

Science

Science