Our service for Transposase Accessible Chromatin with high-throughput single cell sequencing (ATAC-seq) is a technique designed to identify open chromatin regions within the genome at single cell resolution.
Our End-to-End service offers comprehensive support throughout the entire ATAC-seq workflow from tissue dissociation to bioinformatics analysis, ensuring personalized assistance wherever needed.
This method employs a hyperactive Tn5-transposase to uncover the accessible regions of a genome, thereby facilitating the understanding of regulatory landscapes by pinpointing areas where the chromatin is not tightly packed. Through the integration of high-throughput sequencing, ATAC-seq provides a comprehensive map of chromatin accessibility, offering insights into gene regulation mechanisms and cellular function. Notably, advancements in this technology have led to the development of single-cell ATAC-seq (scATAC-seq), tailored for dissecting cell-type-specific chromatin accessibility within heterogeneous tissue samples. This refinement is particularly vital in the context of single-cell analyses, where traditional scRNA-seq methodologies may falter due to their inherent technical constraints, such as the underdetection of low-abundance transcripts including transcription factors (TFs), potentially resulting in misleading negatives. (Ranzoni AM et al., Integrative Single-Cell RNA-Seq and ATAC-Seq Analysis of Human Developmental Hematopoiesis. Cell Stem Cell. 2021 doi: 10.1016/j.stem.2020.11.015)
The scATAC-seq service offered by Singleron utilizes our SCOPE-chip technology to enable precise and sensitive genome-wide mapping of chromatin accessibility.
The ATAC-Seq process begins with the exposure of genomic DNA to Tn5, a transposase known for its robust activity. Tn5 intricately cuts the DNA and preferentially targets areas of open chromatin, simultaneously inserting sequencing adapters in a step known as tagmentation. Following tagmentation, the DNA fragments are loaded onto the chip and captured by uniquely barcoded beads. These barcodes are then associated with the DNA fragments, forming libraries that are amplified through PCR before undergoing sequencing. Additionally, we executed single-cell Assay for Transposase-Accessible Chromatin using sequencing (scATAC-seq) on a tumor in parallel with single-cell RNA sequencing (scRNA-seq) to generate paired datasets elucidating cell-type-specific chromatin accessibility and transcriptional landscapes.
scATAC-seq of mouse embryos yielded 7,880 nuclei from 15,000 cells. Each cell produced ~30,741 reads (58.5% high-quality), with 55.42% overlapping chromatin accessibility peaks, potentially linked to gene regulation.
Precise annotation of single-cell transcriptome is achieved through marker genes in mouse embryo, followed by accurate annotation of single-cell ATAC cell atlas using the “label transferring” method.
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