Rapid preparation of tissue DNA from paraffin-embedded blocks and analysis by polymerase chain reaction.
We have developed a new, easy, and more rapid method for DNA preparation, which avoids contamination. With this method, manual surgical blade scrapings from precisely targeted areas of paraffin block surfaces, without microtome cutting, were used to obtain tissues from 10 different neoplasms. Our results indicate the feasibility of DNA extraction from the scraped paraffin tissue for molecular genetic analysis. We applied this technique successfully to screen for the presence of human papillomavirus using the polymerase chain reaction (PCR) procedure in cases of endocervical, esophageal, and nasopharyngeal carcinomas, and to examine the expression of p53 gene from prostate and gastric adenocarcinomas. We conclude that this procedure is also suitable for purification of PCR products in analysis of the mutation or loss of allelic genes by Bstu I endonuclease digestion.
Department of Pathology, Tulane University School of Medicine, New Orleans, Louisiana.
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Chen
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Clejan
D001483:Base Sequence; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification; D004938:Esophageal Neoplasms / Q000737:chemistry; D005091:Exons; D005240:Feasibility Studies; D005260:Female; D016158:Genes, p53; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009303:Nasopharyngeal Neoplasms / Q000737:chemistry; D009369:Neoplasms / Q000737:chemistry* / Q000473:pathology; D015345:Oligonucleotide Probes / Q000737:chemistry; D027383:Papillomaviridae / Q000302:isolation & purification; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D011471:Prostatic Neoplasms / Q000737:chemistry; D013274:Stomach Neoplasms / Q000737:chemistry; D002583:Uterine Cervical Neoplasms / Q000737:chemistry
D004273:DNA, Neoplasm; D004279:DNA, Viral; D015345:Oligonucleotide Probes