Molecular Approaches to Diagnosis in Ewing Sarcoma : Targeted RNA Sequencing

Molecular testing of pathognomonic gene fusions is mandatory for small round cell tumor diagnosis, including Ewing sarcoma which is indeed defined by a variety of chimeric genes. Reference laboratories are increasingly implementing NGS-based techniques to overcome several limitations of conventional singleplex determinations. We have been early adopters of a targeted-RNA sequencing method based on Anchored multiplex PCR, which allows assessing several fusion transcripts simultaneously with previous knowledge of only one partner gene. Here we describe in detail our protocol and tips for nucleic acid extraction, library preparation, sequencing, and reporting of gene fusions.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:2226

Enthalten in:

Methods in molecular biology (Clifton, N.J.) - 2226(2021) vom: 16., Seite 105-116

Sprache:

Englisch

Beteiligte Personen:

Salguero-Aranda, Carmen [VerfasserIn]
Diaz-Martin, Juan [VerfasserIn]

Links:

Volltext

Themen:

Anchored Multiplex PCR (AMP)
Biomarkers, Tumor
Ewing sarcoma
FFPE sample
Gene fusion
Journal Article
Nucleic acid extraction
Oncogene Proteins, Fusion
Precision diagnostics
Targeted RNA-seq

Anmerkungen:

Date Completed 01.04.2021

Date Revised 01.04.2021

published: Print

Citation Status MEDLINE

doi:

10.1007/978-1-0716-1020-6_8

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM318911884