SarcomaFusion Kit – 24 reactions

The SarcomaFusion is a molecular diagnostic assay designed to prepare sequencing libraries for the detection of fusion transcripts associated with sarcomas. The test uses a ligation-dependent RT-PCR approach based on specific oligonucleotide probe pairs and includes an internal GAPDH control.

By studying 210 genes, SarcomaFusion enables the detection of fusion transcripts associated with more than 80 types of bone and soft tissue tumours. The assay can be performed using total RNA extracted from fresh, frozen or formalin-fixed, paraffin-embedded (FFPE) tumour tissues or biopsies.

The workflow consists of reverse transcription, hybridization of specific oligonucleotide probes, ligation, and PCR amplification with incorporation of molecular barcodes and sequencing adapters. No purification is required before library generation, which limits material loss and supports analysis of samples with potentially degraded RNA, including FFPE specimens.

The resulting sequencing libraries are purified, quantified and sequenced using compatible Illumina MiSeq or NextSeq 500/550 platforms. Generated FASTQ files are analysed using the Genexpath RT-MIS software for demultiplexing, identification and quantification of fusion-associated sequences.

GENEXPATH SarcomaFusion

Product Features

Molecular assay for the detection of sarcoma-associated fusion transcripts
Ligation-dependent RT-PCR technology
Targets 210 genes
Covers fusion transcripts found in more than 80 types of bone and soft tissue tumours
Suitable for fresh, frozen and FFPE tumour samples
Designed for total RNA extracted from sarcoma tumour tissues or biopsies
Requires 50–500 ng of RNA per sample
Short target sequences of approximately 40–60 bases support analysis of potentially degraded RNA
No purification required before sequencing library generation
Includes GAPDH internal positive control
Molecular barcodes enable multiplexing of samples in the same sequencing FlowCell
Compatible with Illumina MiSeq and NextSeq 500/550 sequencing platforms
FASTQ data analysed using RT-MIS software
Generates analysis reports indicating the presence or absence of fusion transcripts
For in vitro diagnostic use
For professional use only

Product Specifications

Catalog Number	GEP-SF24
Product Type	Sarcoma fusion transcript detection assay
Input Material	Total RNA
Recommended RNA Input	50–500 ng in 2.5 µL
Internal Control	GAPDH internal positive control
Kit Format	24 reactions
Probe Mix	GEP-SFPM, 54 µL
SarcomaFusion Barcodes	12 GEP-BC barcodes, 9 µL per barcode
Sequencing Primer	GEP-SP-001, 144 µL
GAPDH Barcodes	12 GEP-BCC barcodes, 9 µL per barcode
GAPDH Sequencing Primer	GEP-SP-002, 144 µL
Sequencing Platforms	Illumina MiSeq and NextSeq 500/550
Data Format	FASTQ
Analysis Software	Genexpath RT-MIS
Storage	−25°C to −15°C
Shelf Life	12 months from the production date
Intended Use	In vitro diagnostic use; professional use only

SarcomaFusion Workflow

Reverse transcription: Total RNA is converted into cDNA using a reverse transcription reaction.
Probe hybridization: Specific oligonucleotide probes hybridize to the target sequences.
Ligation: Hybridized probe pairs are ligated to generate products corresponding to the targeted fusion sequences.
Amplification and barcoding: Ligation products are amplified by PCR while incorporating sample-specific molecular barcodes and sequencing adapters.
Purification: Amplified sequencing libraries are purified using AMPure XP magnetic beads.
Library quantification: Purified libraries are quantified by fluorimetry using the Qubit dsDNA HS assay.
Pooling and sequencing: Libraries are diluted, pooled and sequenced on compatible Illumina MiSeq or NextSeq 500/550 platforms.
Data analysis: FASTQ files are analysed using Genexpath RT-MIS for demultiplexing, identification and quantification of fusion-associated sequences.

Product Details

Reference	Description
GEP-SF24	SarcomaFusion Kit – 24 reactions