Single-cell Full-length Transcriptome Sequencing

Single-cell sequencing technology provides many new discovery perspectives for basic scientific research, clinical diagnosis, drug development and other fields. The current mainstream single-cell sequencing is mostly based on 10X Genomics, BD Rhapsody and other single-cell capture equipment to obtain cDNA, then interrupt, amplify, build a library, and use next-generation sequencing to quantify the overall  genes. However, genes use different transcripts of mRNA in different tissues and different cell subpopulations, including IncRNA; in addition, structural variations such as SNVs and fusion genes are also tissue and cell specific, currently based on single-cell data from next-generation sequencing. It is limited to 100~150bp at the 3' or 5' end, so it is difficult to meet such requirements. Although traditional Smart-seq can achieve full-length transcript coverage, transcript structure analysis requires assembly and low cell throughput with high cost, makes it difficult to study alternative splicing at the single-cell level.

Technologies such as Pacbio and Nanopore can solve this problem with their advantages of long read length. Therefore, if the next generation sequencing and long-read sequencing can be combined, the full-length sequence of the mRNA can be obtained, and the cell subgroup can be located through the Cell Barcode information, which can overcome the problem in the field of single-cell research. Mining valuable specific transcripts from the huge second-generation + third-generation single-cell data, that is, genes + transcripts, can bring more valuable information for single-cell clinical transformation and drug target discovery.

What We Offer

CD Genomics can provide solutions from sample preservation, transportation, single cell suspension preparation, to single cell sorting, library building and data analysis based on more than ten years of experience in single cell genomics services.

Meanwhile, around the single cell enrichment and detection platform, single cell sequencing technology platform and single cell data analysis platform based on AI algorithm, we have established a variety of unique analysis processes and methods such as single cell transcriptome, spatial transcriptome, integrated single cell multi-omics, etc. We are especially good at analysis projects such as classification, functional analysis, cell interactions and drug target screening of various types of immune cells and stromal cells. We have accumulated hundreds of single cell analysis methods and millions of single cell databases to provide professional R&D services for single cell clinical translation projects.

CD Genomics officially launches single-cell full-length transcript sequencing service, i.e., transcriptional and genetic variation studies at the single-cell cDNA level. Through one capture and two library building, single cell clustering and transcript information can be obtained at the same time

Single-cell Full-length Transcriptome Sequencing Workflow

Currently, the technology offers potential solutions to the following research problems:

1. Discover the mutations carried by different cells and discover the rules of gene expression

2. Mining functional genes, such as membrane proteins, secreted proteins, transcription factors, and other transcript usage, and discovering transcripts with new functions;

3. Discover the cell subpopulation where the fusion gene is, and study their relationship with other tumor cells;

4. Discover new subgroup-specific lncRNA

5. Obtaining subgroup-specifically expressed transcripts can assist small nucleic acid drug development companies to design siRNA interference fragments for the specific transcripts and improve the effectiveness of small nucleic acid interference targets.

Single-cell Full-length Transcriptome Sequencing Analysis

Sample Requirements

Sample type: fresh tissue, primary cells, cell lines, etc.

Sample source: blood extraction, magnetic bead enrichment, flow cytometry enrichment, tissue dissociation, etc.

Sample size and other quality control requirements:

(1) Cell suspension: >10*number of target cells (at least 10,000 cells);

Survival rate>85%

Concentration 500-1,000 cells/ul;

No adhesion between cells (agglomeration rate <5%)

No cell debris or other particles larger than 40um

Reverse transcription inhibitors and non-cellular nucleic acid molecules are absent.

(2) Blood: whole blood anticoagulated with EDTA (no heparin anticoagulated), >5ml.

(3) Tissue: 4-5 pieces of fresh tissue of 0.3cm×0.3cm (no more than 0.5×0.5cm).

The number of captured cells and the amount of sequencing data:

* Whole exome sequencing: 20G