Adapter Template Oligo-Mediated Sequencing.
A Revolutionary New DNA-Capture Method for Next Generation Sequencing
ATOM-Seq Technological Overview
XCeloSeq library preparation products employ the novel, patented, GeneFirst breakthrough technology, ATOM-Seq. This approach uses a simple and elegant chemistry to capture DNA, where original molecules act as primers and their 3’ ends are extended by a polymerase.
Using the ATOM-Seq approach, Unique Molecular Identifiers (UMIs) and universal adapter sequences are incorporated directly into available 3' ends of every original sample molecule.
Sample DNA or cDNA is combined with the Adapter Template Oligo
The DNA molecules anneal to the 3’ end of the ATO.
Hands-on Time: 5 min
Incubation Time: 70 min
After annealing, a polymerase extends the original molecule, using the ATO sequence as a template.
Using its ATO template, the polymerase incorporates both a Unique Molecular Identifier (UMI) and a universal priming site into the captured molecule.
Hands-on Time: 5 min
Incubation Time: 35 min
A universal primer is used for multiple rounds of linear amplification, improving error correction. This also means that all original copies can be represented in downstream reactions, even when the linear product is divided between sense- and antisense-strand targeted enrichment
Protocol time includes all steps shown, from initial ATO Reaction to final, bead-purified library.
Total: 5h 40m
Hands-on: 1h 20m
Allele frequency: ≥0.1%
Total: 7h 20m
Hands-on: 1h 55m
Both known and
unknown gene fusions
Total: 4h 45m
ATOM-Seq Key Advantages
This unique approach is ligation free and is set apart from conventional PCR-based approaches by requiring only a single target-specific primer for enrichment. This offers numerous advantages that make XCeloSeq particularly well suited to challenging clinical material such as cell-free DNA (cfDNA) from liquid biopsies or FFPE-preserved RNA.
Green ticks indicate aspects the technologies can do.
Green bars indicate aspects of the technologies which are inefficient.
Red crosses indicate aspects the technologies cannot or have not been optimised to do.