Single-Cell Sequencing

Single-Cell Sequencing

Single cell sequencing is a new technology for high-throughput sequencing analysis of genetic material at individual cell level, which enables comprehension of the complex genetic variability and structural and functional properties of individual cells. Genome, transcriptome, epigenome, and proteome sequencing are fundamental constitutions that reflect single-cell developmental trajectory. The main process of single cell sequencing includes single cell isolation, single cell lysis, nucleic acid amplification, high-throughput sequencing, data processing and data analysis. The utilization of microfluidics technology in single-cell sequencing process improves the throughput and sensitivity of cellular analysis and reduces the risk of contamination and bias during the amplification process.

Microfluidics Applied in Single-Cell Sequencing

Single cell isolation

Microfluidic devices enable isolation of individual cells using the following principles: droplet-based isolation, valve-based isolation, trap-based isolation, microwell-based isolation, DMF-based isolation, dielectrophoresis-based isolation.

Single cell lysis

Microfluidics-based lysis minimizes lysate dilution, thus increasing assay sensitivity to a large extent. The common cell lysis methods used in microfluidics devices include physical, chemical and enzymatic methods.

• Physical cell lysis: Mechanical, thermal and electrical
• Chemical cell lysis: Surfactants, such as sodium dodecyl sulfate (SDS) and Triton X-100
• Enzymatic cell lysis: Proteinase, such as pepsin and trypsin

Nucleic acid amplification

There are two common amplification methods: Whole-genome amplification (WGA) and whole-transcriptome amplification (WTA).

Methods for WGA: Degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), multiple displacement amplification (MDA), and multiple annealing and looping-based amplification cycles (MALBAC).

Methods for WTA: Traditional PCR, modified PCR, T7-in vitro transcription (IVT) and Phi29 DNA polymerase-mediated RNA amplification.

Single-cell sequencing

• Single-Cell Genome Sequencing (scDNA-Seq)

Single-cell genome sequencing reveals intercellular variation and heterogeneity of the genome, such as single-nucleotide variants (SNVs) and copy number aberrations (CNAs).

• Single-Cell Epigenome Sequencing

Single-cell epigenome sequencing studies DNA modifications, histone modifications, DNA accessibility, and chromatin conformation.

• Single-Cell Transcriptome Sequencing (scRNA-Seq)

Single-cell transcriptome sequencing measures gene expression at the single cell resolution, identifying biologically relevant differences in cell clusters.

• Single-Cell Proteome Sequencing

Single-cell proteomics can reveal the cellular phenotypic heterogeneity and cell-specific functional networks of biological processes.

For more information about our single-cell sequencing service, please contact us.