Robust Detection of Gene Amplification in Formalin-Fixed Paraffin-Embedded Samples by Fluorescence in situ Hybridization.
J Vis Exp
; (209)2024 Jul 12.
Article
in 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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Gene Amplification
/
Paraffin Embedding
/
In Situ Hybridization, Fluorescence
/
Formaldehyde
Limits:
Humans
Language:
En
Journal:
J Vis Exp
Year:
2024
Document type:
Article
Country of publication:
Estados Unidos