XCeloSeq Targeted RNA Enrichment Kits
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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
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
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Both known and
unknown gene fusions
Detects:
Input Amount*:
Recommended:
5-200ng
Minimum: 1ng
Protocol Time:
Total: <7.5 hours
Hands-on: 2 hours
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 cDNA, 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 protocol document, available here.
Please also see the relevant product insert(s) for kit-specific information and protocol steps, given below.
Please note that an XCeloSeq UDI Kit (containing unique premixed i5 and i7 indexing primers) must be purchased separately to use any of the library preparation kits listed on this page. Please see below for more information.
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XCeloSeq Unique Dual Indexing Sets (IDX1-01 to IDX1-12)
The XCeloSeq Unique Dual Indexing (UDI) Sets allow for multiplexing of up to 96 different samples each with a unique combination of i5 and i7 indexes for next-generation sequencing on Illumina sequencing platforms. All index oligos undergo rigorous quality control procedures including generation and sequencing of indexed libraries to ensure high quality performance with all compatible XCeloSeq Library Preparation Kits.
When purchasing UDI Sets for use in combination with any of the XCeloSeq Targeted RNA Enrichment Kits up to 96 samples can be multiplexed together when purchasing all 12 Sets. Each UDI combination contains enough for 8 samples, for a total of 128 samples for every UDI Set.
At least one UDI Set must be purchased with XCeloSeq Targeted RNA Enrichment Kits.
Click Here to download product information for ordering, multiplexing, and sample requirements.
*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.
For more information or for sales enquiries, please contact us here or at info@genefirst.com