Accel-NGS® Adaptase® Module for Single-Cell Methyl-Seq
Explore the Methylome at Single Cell Resolution
The Accel-NGS Adaptase Module constructs NGS libraries from bisulfite-converted DNA from single cells. The Adaptase Module maximizes the recovery of low concentrations of single stranded DNA and libraries consistently exhibit superior complexity with reduced composition bias to provide a more faithful representation of the methylome. Using the in-line barcoding option, cells can be mulitplexed to increase throughput and further reduce library prep costs.
The Adaptase Module can be used for many applications, such as cataloging cell populations within heterogeneous tissues, assessing normal tissue for regulation of cellular mechanisms (e.g., differentiation), gaining insight into epigenetic alterations in disease states, and exploring across species to identify evolutionary conservation of epigenomic regulation.
- High mapping rate
- >400 bp library insert size improves data output
- Workflow uses a single random priming cycle
- Greater data output per run and reduced sequencing cost
- Scalable cell and library indexing options for high throughput sequencing
- Easier low bias workflow
Following lysis and bisulfite conversion of single cells, random priming of bisulfite-converted DNA incorporates a truncated P5 adapter (Truncated Adapter 1) to 5’ ends of synthesized fragments, and also effectively reduces fragment size to ~400 bp for suitability with Illumina® sequencing. Optionally, indexed random primers can be used for downstream multiplexing capability. Unused reagents are digested followed by a purification step. Next, the highly efficient Adaptase step simultaneously tails and ligates the truncated P7 adapter (Truncated Adapter 2) to the 3’ end of single-stranded products, which completes library construction. Library amplification is then performed using custom primers that incorporate dual indexing.
The Accel-NGS Adaptase Module requires commercially-available oligonucleotide primers, enzymes, and buffers (ordered separately) for complete functionality. Please see the Accel-NGS Adaptase Module for Single-Cell Methyl-Seq Library Preparation Protocol for a complete list of required third-party materials not supplied by Swift Biosciences.