Single-cell RNA Sequencing
Unprecedented insights into cellular diversity and function.
What is Single-cell RNA Sequencing?
Single-cell sequencing is a cutting-edge technology that allows researchers to analyze individual cells’ genetic and molecular characteristics, providing unprecedented insights into cellular diversity and function.
Unlike traditional bulk sequencing, which averages signals from thousands of cells and masks the differences between individual cells, single-cell sequencing reveals the heterogeneity within a population, enabling researchers to identify distinct cell subpopulations, understand cell-specific gene expression patterns, and observe how individual cells contribute to overall tissue function or disease progression.
Applications of Single-cell RNA Sequencing
The field of single-cell genomics is advancing rapidly, providing new insights into complex biological systems—from the genomics of human cancer to the diversity of microbial ecosystems.
Common applications of single-cell sequencing include cancer research, cardiovascular research, developmental biology, drug discovery, infectious disease, immunology, neuroscience, Plant and Agricultural Research ,and stem cell research.
Single-cell Sequencing Workflow with DNBSEQ™ Technology
DNBSEQ sequencing technology is compatible with a range of single-cell library preparation and analysis solutions, including those from 10x Genomics, Parse, and Fluent Biosciences. Unlike traditional PCR amplification, DNBSEQ uses rolling circle replication (RCR), which ensures highly accurate sequencing with exceptional single-base resolution. This advanced approach eliminates common issues such as index hopping and reduces duplicate levels, leading to more reliable and precise results.
Single-cell sequencing can be cost-intensive due to the large number of cells that need to be sequenced. The DNBSEQ-G400 and DNBSEQ-T7 sequencing platforms address this challenge by offering higher accuracy, exceptional flexibility, and significantly improved cost efficiency, making them ideal solutions for single-cell applications.
Sample to Analysis Workflow
Sample Prep
Parse Biosciences:
Evercode Cell
Fixation v2 Kit
Single-cell Library Prep
10x Genomics:
Chromium Single Cell
Parse Biosciences:
Evercode WT Mini
Evercode WT
Evercode WT Mega
Fluent Biosciences:
PIPseqV
Library Conversion
DNBSEQ OneStep Library Conversion Kit
Analysis
Parse Biosciences:
Data Analysis Software
Sample Prep
Prepare Cell Suspension
Single-cell Library Prep
PIPseq V5 T20 3′
Single Cell RNA Kit
Library Conversion
DNBSEQ OneStep Library Conversion Kit
Analysis
PIPseeker Analysis
Single-cell Sequencing Solution by Parse and DNBSEQ-G400
The study compared the sequencing performance of Complete Genomics’ DNBSEQ-G400 and Illumina’s NovaSeq 6000 using single-cell RNA libraries. The cancer cell lines and single-cell RNA libraries were provided and prepared by Vevo Therapeutics (Vevo) using Parse Biosciences kits. In summary, the DNBSEQ-G400 delivered comparable or slightly superior results in single-cell sequencing compared to the NovaSeq 6000.
The UMAP visualization of Vevo’s scRNA-Seq data (the figure on the right), with cells color-coded based on sequencing technology: DNBSEQ-G400 (red) and NovaSeq 6000 (blue) demonstrated the high degree of similarity between the samples sequenced on the different platforms.
Single-cell Sequencing Solution by Fluent PIPseq and DNBSEQ-G400
The DNBSEQ-G400 sequencing platform is an excellent choice for routine single-cell RNA-seq analysis using the Fluent Biosciences PIPseq kit. With high sequencing quality (>93% Q30), full compatibility with the PIPseeker analysis pipeline, and performance that matches or exceeds the NextSeq 2000, the DNBSEQ-G400 delivers reliable and precise results. Moreover, the cost of generating PIPseq data with this system is estimated at just $0.11 per cell, offering an affordable solution for single-cell studies.
