Single-Cell Full-Length Transcriptome Sequencing

Single-Cell Full-Length Transcriptome Sequencing

Single-cell full-length transcriptome sequencing (RNA-seq) is a technology that uses the 10x Genomics Chromium (10x) platform to obtain single-cell cDNA and combine it with transcriptome analysis on a long-sequence sequencing platform. This method combines the advantages of full-length transcriptome and single-cell sequencing, providing a powerful tool for discovering and quantifying isoforms, analyzing alternative splicing at the single-cell level.

Overview of Single-Cell RNA-Seq with Long-Read Sequencing

Single-cell RNA sequencing (scRNA-seq) has made up for the limitations of bulk RNA sequencing (RNA-seq). This method can resolve gene expression profiles in single cells for more accurate transcriptome assessment, and is currently receiving widespread attention and application. There are two main characteristics of the single-cell experiments, including the barcoding of cDNA synthesized by each cell with cell-specific barcodes and of individual mRNA molecules with molecular barcodes. Among them, the former method allows better quantification of transcripts at the single-cell level. Since molecular barcodes are usually degraded, it requires the high accuracy of the sequencing platform. ScRNA-seq is now a widely adopted approach for profiling transcriptomic heterogeneity. However, assessing transcript-level changes between cell types using current scRNA-seq protocols remains challenging, as they rely on short-read sequencing. Nowadays, long-read sequencing technology is widely used in scRNA-seq to generate complete transcript information in a single cell, overcoming the fundamental limitations of short-read sequencing.

Our Service Can Help Researchers Achieve

Workflow of Our Services

10X Genomics captures 500-10,000 cells in a single sequencing run with extremely high cell throughput. Cells are labeled by 10X Genomics in a water-in-oil state, reverse transcribed into cDNA, then ligated by head-to-tail ligation process of the cDNAs and constructed into a long-read sequencing library of 10-15kb in length, which is then sequenced.

Workflow of Our Services-CD Genomics.

Analysis Pipeline and Contents

Analysis Pipeline and Contents-CD Genomics.

Features and Benefits of Our Services

At CD Genomics, we offer two strategies for our customers to choose from, including Pacific Biosciences (PacBio) and Oxford Nanopore Technologies (ONT) single-cell full-length transcriptome sequencing. Both methods have their own advantages in the field of RNA-Seq. Based on advanced platforms and years of experience in transcriptome analysis, we are committed to providing the best strategy to meet the project needs of our global clients. Our single-cell full-length transcriptome sequencing service has many applications. In addition to normal developmental and physiological studies, we focus on disease studies, such as tumor research and viral infections. If you have additional requirements or questions about our services, please feel free to contact us.

References

  1. Oikonomopoulos, S., et al. (2020). "Methodologies for transcript profiling using long-read technologies." Frontiers in genetics, 606.
  2. Tian, L., et al. (2021). "Comprehensive characterization of single-cell full-length isoforms in human and mouse with long-read sequencing." Genome biology, 22(1), 1-24.
  3. Wang, Q., et al. (2021). "Single-cell transcriptome sequencing on the Nanopore platform with ScNapBar." RNA, 27(7), 763-770.
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