Robust Detection of Gene Amplification in Formalin-Fixed Paraffin-Embedded Samples by Fluorescence in situ Hybridization.
J Vis Exp
; (209)2024 Jul 12.
Article
em En
| MEDLINE
| ID: mdl-39072619
ABSTRACT
Focal gene amplification, such as extrachromosomal DNA (ecDNA), plays an important role in cancer development and therapy resistance. While sequencing-based methodologies enable an unbiased identification of ecDNA, cytogenetic-based techniques, such as fluorescence in situ hybridization (FISH), remain time and cost-effective for identifying ecDNA in clinical specimens. The application of FISH in formalin-fixed paraffin-embedded (FFPE) tissue samples offers a unique avenue for detecting amplified genes, particularly when viable specimens are not available for karyotype examination. However, there is a lack of consensus procedures for this technique. This protocol provides comprehensive, fully optimized, step-by-step instructions for conducting FISH to detect gene amplification, including ecDNA, in FFPE tissue samples which present unique challenges that this protocol aims to overcome and standardize. By following this protocol, researchers can reproducibly acquire high-quality imaging data to assess gene amplification.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Amplificação de Genes
/
Inclusão em Parafina
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Hibridização in Situ Fluorescente
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Formaldeído
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article