XCeloSeq® Targeted RNA Enrichment Kits
NGS Library Preparation kits for cancer detection from FFPE RNA samples
The XCeloSeq Targeted RNA Enrichment Kits generate high quality sequencing libraries for detection of cancer-related gene fusion events from FFPE-preserved material.
ENHANCED LIBRARY COMPLEXITY
All XCeloSeq products use the ATOM-Seq® capture chemistry which is built to be uniquely suited for use with FFPE material and liquid biopsy, delivering unrivaled library complexity
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FUSION-AGNOSTIC TARGETED ENRICHMENT
XCeloSeq target enrichment uses a single target-specific primer in the conserved exon of fusion events, allowing for detection of both known and unknown fusion partners
STREAMLINED AND FAST WITH HIGHEST SAMPLE RETENTION
XCeloSeq protocols have been designed to be completed within 1 day with the fewest bead purifications and shortest hands-on times
Both known and
unknown gene fusions
Detects:
Protocol Time:
Total: 7h 20m
Hands-on: 1h 55m
Input Amount*:
Recommended: 5-200ng
Minimum: 1ng
The XCeloSeq Targeted RNA Workflow
Using the ATOM-Seq DNA Capture Method, Unique Molecular Identifiers (UMIs) and universal adapter sequences are incorporated directly into available 3' ends of sample cDNA . Then, two rounds of target-specific PCR generate sequencing-ready libraries.
Double Strand cDNA Synthesis
Hands-on Time: 35 minutes
Incubation Time: 1 hour
Sample RNA is copied to produce double stranded RNA, allowing interrogation of gene fusions in either orientation.
ATOM-Seq DNA Capture​
Hands-on Time: 10 minutes
Incubation Time: 1 hour and 45 minutes
cDNA is combined with the Adapter Template Oligo (ATO).
Incorporation of universal priming site and Unique Molecular Indicator directly onto the sample cDNA molecule.
Two rounds of Nested,
Target-Specific PCR
Hands-on Time: 1 hour and 10 minutes
Incubation Time: 2 hour and 35 minutes
Multiplexed target-specific primers are paired against a universal primer for target enrichment PCR. Using only a single target-specific primer means that both known and novel fusion partners can be enriched for in this way.
A second round of target-specific PCR using a nested primer maximises on-target rate of sequencing, and incorporates dual-index sample barcodes (i5 and i7).
Sequencing-ready library in 7 hours, 20 minutes.
The XCeloSeq Targeted RNA Enrichment Product Range:
All Targeted RNA Enrichment Kits use a common Instructions For Use document, available here.
Please also see the relevant product insert(s) for kit-specific information and protocol steps, given below.
XCeloSeq Fusion Research Kit
A comprehensive panel for detection of known and unknown gene fusion partners for a variety of cancer types, from FFPE-preserved samples.
Product code: SEQ007
Gene Targets: 74
Reactions per kit: 8
View Product Insert
Targeting Primers: 444
View target genes
XCeloSeq Lung Cancer Fusion Kit
A panel for detection of known and unknown gene fusion partners in lung cancer, from FFPE-preserved samples.
Product code: SEQ008
Gene Targets: 15
View Product Insert
Reactions per kit: 8
Targeting Primers: 158
View target genes
XCeloSeq Solid Cancer Fusion Kit
A panel for detection of known and unknown gene fusion partners in solid cancers, from FFPE-preserved samples.
Product code: SEQ012
Gene Targets: 53
Reactions per kit: 8
View Product Insert
Targeting Primers: 350
View target genes
XCeloSeq Indexing Kits (SEQ005)
Dual indexing primers to enable sample multiplexing. Each kit contains either eight i5 or eight i7 sample index primers.
Kits can be purchased separately to suit your multiplexing requirements.
These primers can be used as substitutes for the default pair of sample indexing primers included in any XCeloSeq Library preparation kit, including XCeloSeq Targeted RNA kits.
*Higher quantities within this range will improve maximum sensitivity, recommended maximum is 200 ng total FFPE RNA or 100 ng high quality RNA. The product supports capture with down to 1.0 ng of RNA, however this is not recommended as it will lead to reduced sensitivity. Cell-free total nucleic acids and cell-free RNA are also supported as alternative starting materials, when using these materials maximising starting material within the above range will help ensure high quality results.
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