Promoter methylation analysis on microdissected paraffin-embedded tissues using bisulfite treatment and PCR-SSCP

Détails

ID Serval
serval:BIB_22AAE54DFB3C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Promoter methylation analysis on microdissected paraffin-embedded tissues using bisulfite treatment and PCR-SSCP
Périodique
BioTechniques
Auteur⸱e⸱s
Bian  Y. S., Yan  P., Osterheld  M. C., Fontolliet  C., Benhattar  J.
ISSN
0736-6205 (Print)
Statut éditorial
Publié
Date de publication
2001
Volume
30
Numéro
1
Pages
66-72
Notes
PT - Research Support, Non-U.S. Gov't PT - Technical Report
Résumé
Methylation-sensitive single-strand conformation analysis (MS-SSCA) is a new method of screening for DNA methylation changes. The combination of bisulfite modification and PCR results in the conversion of unmethylated cytosines to thymines, whereas methylated cytosines remain unchanged. This sequence conversion can lead to methylation-dependent alterations of single-strand conformation, which can be detected by SSCA. An analysis of mixtures of methylated and unmethylated DNA at known ratios revealed that the relative intensities of the corresponding bands following MS-SSCA were maintained. MS-SSCA was applied for methylation analysis of human p16 promoter region using genomic DNA obtained from either frozen, fixed, or microdissected fixed tissue sections. MS-SSCA is a rapid, specific, and semiquantitative approach that allows the detection of methylation of the p16 gene promoter. In reconstruction experiments, the method permits the detection of 10% or less of cells harboring a methylated p16 promoter. We have been successful in analyzing by MS-SSCA almost all (96%) tumor samples microdissected from archival paraffin-embedded fixed tissue sections and obtaining reproducible results. In addition, when microdissection was performed, the clonality of this genetic alteration could be identified
Mots-clé
Dna/genetics/metabolism/DNA Methylation/Dissection/Female/Humans/Neoplasms/Pathology/Paraffin Embedding/Placenta/Polymerase Chain Reaction/methods/Polymorphism,Single-Stranded Conformational/Promoter Regions (Genetics)/Reproducibility of Results/Sensitivity and Specificity/Sulfites/Tissue Fixation/Tumor Cells,Cultured
Pubmed
Web of science
Création de la notice
29/01/2008 18:35
Dernière modification de la notice
20/08/2019 13:00
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