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Single Cell Sequencing Service
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Spatial transcriptome sequencing

Platform Introduction

Wuhan Biological Sample Bank (Wuhan National Human Genetic Resources Bank) is one of the earliest units engaged in the research and development and service of single-cell sequencing technology in China. It first introduced 10x Genomics Chromium equipment and technology platform. At present, it has successfully carried out single-cell sequencing of human, mouse, fish, poplar and other multi-species and tissues (including tumor, blood, kidney, liver, intestine, bone, plant protoplast, etc.). In April 2021, it will become a single-cell sequencing co-construction laboratory in central China officially certified by 10x Genomics.

平台简介

 

At the same time, the company has professional cell sorting equipment and rich cell sorting experience; It has a high-throughput sequencing equipment (Illumina Novaseq) in Central China and Huada DNBSEQ-T7; It has a server cluster with strong computing capability and an independent online analysis platform; It can provide customers with all-round and full-process single-cell research solutions.

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Service Items

Single cell sequencing
Spatial transcriptome sequencing
Single Cell Multiome ATAC + Gene Expression
Single Cell Multiome ATAC + Gene Expression combines ATAC with transcriptome sequencing to identify complex cell populations through integrated analysis of epigenomic and gene expression profiles and detect cellular heterogeneity to uncover hidden information. Combining the discovery of regulatory elements with gene expression could help to explore the gene regulatory interactions that drive cell differentiation, development and disease. Sample types: animal tissues, single cell suspensions, nuclear suspensions, cell lines.   Experimental process 单细胞多组学ATAC+基因表达实验流程图 Analysis process Flow Chart of ATAC+Gene Expression Analysis in Single Cell Multiomics 　　 Application directions a. Biomarker discovery; b. Cell lineage and developmental program tracking; c. Cell heterogeneity and detection of rare cell populations; d. Gene regulatory networks; e. Response to therapeutic interventions.   Product advantages a. Efficiently distributing 500-10,000 nuclei per lane, analyzing up to 80,000 nuclei per run ; b. Extensible to run up to 8 samples in parallel; c. Nuclei recovery up to 65%; d. High sensitivity; e. Low microfluidic double rate (less than 1% per 1,000 nuclei); f. Suitable for studies with cell lines, primary cells, fresh and frozen samples.

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Single Cell ATAC
Single-cell ATAC-seq is to study chromatin accessibility at the single-cell level and is mainly used for the detection of transcriptional regulatory sequences, which can be combined with single-cell RNA-seq to analyse all actively transcribed regulatory sequences in the genome. This technique has been used in many research areas such as tumor heterogeneity studies, gene regulatory network analysis, cell lineage tracing, biomarker discovery, etc. ATAC-Seq (Assay for Transposase Accessible Chromatin with high-throughput sequencing) uses Tn5 transposase to cleave nuclear chromatin regions which is open at a specific space-time and then adds primers for high-throughput sequencing to obtain the regulatory sequences of all active transcripts in the genome at that specific space-time condition.   ATAC-Seq schematic diagram   The single cell ATAC-Seq technology relies on the microfluidic technology of the 10x Genomics platform, combining characteristics of different cells of sequence tags distinguished population with ATAC-Seq technology to build chromatin accessibility profiles for thousands of single cells. Sample types: cell lines, primary cells, fresh and frozen tissues, etc. Schematic diagram of 10x Genomics single-cell ATAC-Seq   Sample type Cell lines, primary cells, fresh and frozen tissues, etc. Experimental process 10x Genomics单细胞ATAC-Seq实验流程   Analysis process Flow chart of single-cell ATAC analysis 　　 Application directions a. Studying cell heterogeneity; b. Exploring biomarkers; c. Defining cell types; d. Building gene regulatory networks.   Product advantages a. Detecting open chromatin in transcriptional regulatory regions of single cells; b. Each channel can hold 500~10000 nuclei; c. Nuclei capture rate up to 65%; d. Suitable for the study of cell lines, primary cells, fresh and frozen samples; e. Using the official 10x software, single cell gene regulation can be deeply analyzed.

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Single Cell Immune Profiling
In 2015, 10x Genomics released the Chromium Single Cell System platform based on microfluidic and oil-drop wrapping technologies, enabling high-throughput single-cell transcriptome and single-cell V(D)J sequencing. This technology not only allows perfect matching of TCR/BCR duplexes to obtain TCR/BCR duplex composition, but also could be detailed into the single cell level while obtaining 5' end transcriptome information. New released Single Cell Immunoprofiling of 10x Genomics uses microfluidic chip to prepare single cell systems, selecting universal primers for the 5' end-joins and nested primers for the immune molecular constancy region for V(D)J enrichment to enable full-length sequencing of paired heavy and light chains (B cells) or alpha and beta chains (T cells) at the single cell level. Full-length sequencing is performed, thus providing an efficient technical platform for comprehensive and systematic studies of immunomic libraries, and is important for the study of the molecular mechanisms of disease onset and development of disease. Sample types Animal tissues, other body fluids such as blood, single cell suspensions, cell lines, etc.   Experimental process Experimental flow of 10x Genomics single cell immune group library Analysis process Flow Chart of Bioinformatics Analysis of Single Cell VDJ Immunogroup Bank   Application directions a. Identification of antibodies and discovery of new antibodies; b. Revealing the development of Ig libraries and mapping global Ig libraries; c. Studying infectious diseases, such as the evolutionary spectrum of antibodies in the development of infections; d. Studying the propensity for V(D)J recombination in immune disorders and to search for biomarkers of immune disorders; e. Early detection of cancer, and studying markers for the diagnosis of cancer progression and recurrence, and studying tumor immune mechanisms. Product advantages a. Identification and characterization of rare cell types and biomarkers; b. Analysis of tissue micro environment, disease progression and drug immune response; c. Large-scale antibody and TCR discovery against neoantigens d. Characterizing the immune response to infection by measuring immune cell phenotypes; e. Reduce single cell sequencing costs through large-scale single-cell V(D)J+ expression profiling sequencing; f. Provide flow cytometry sorting services to enable sorting for immune cells with purposes.

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Single Cell Gene Expression
Single cell RNA sequencing (scRNA-seq) is a new technology for high throughput sequencing of mRNA at the single cell level. The principle is to dissociate the sample tissue into  single cell suspension and then carry out high-throughput sequencing of the minute amounts of  mRNA in single cells after efficient amplification. Single cell transcriptome sequencing can solve the problem of cell heterogeneity that is obscured by conventional RNA-seq and help to discover new cell types, which is important for neurobiology research, immunology research and developmental biology research. Sample types: animal and plant tissue, blood and other body fluids, cell lines, single cell suspensions, plant protoplast suspensions,  animal and plant cell nucleus suspensions and so on.   Experimental process 　　10x Genomics单细胞转录组实验流程   Analysis process Flow Chart of Single-cell Transcriptome Bioinformatics Analysis 　　 Application directions a.Large-scale cell atlas construction; b. Identification of rare cell types & cell subpopulation refinement; c. Tumor heterogeneity and tumor microenvironment research; d. Immunological research; e. Cell development and differentiation research. 　　 Product advantages a. Real single cell level sequencing with high cell capture efficiency; b. The team is experienced in sample dissociation and could accept a wide range of sample types; c. One-stop service for sample processing, sorting, loading, sequencing and analysis; d. Short project period and wide range of technical applications.

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Spatial Gene Expression for FFPE
Considering traditional spatial transcriptome sequencing is limited to fresh frozen tissue sections, 10X Genomics introduces the new Visium spatial gene expression solution for FFPE samples (formalin-fixed paraffin-embedded), which breaks the barriers to obtain spatial gene expression information from FFPE tissue samples and get highly sensitive and specific expression and spatial location information of relevant genes from FFPE samples. FFPE sample gene expression is a pioneering solution that complements traditional pathology-driven analysis, enabling true discovery in a morphological context and further expanding the possibilities of spatial biology research.

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Spatial Gene Expression for Fresh Frozen
Principle of the technique When a frozen tissue slice is placed in the capture region of a Visium chip with spatial barcode and has been HE stained and imaged, the tissue is permeabilised and the intracellular mRNA is released and captured by a probe with oligo-dT on the chip. The captured mRNA starts reverse transcription and the resulting cDNA contains spatial barcode sequences. After constructing a library and sequencing, the mRNA transcribed sequences can be mapped to their original location in the tissue, thus obtaining information about the location of gene expression. The core of the Visim spatial transcriptome is the microarray: the formal library is constructed with four capture regions, each containing approximately 5,000 spots, each capable of capturing 1-10 cells, with a distance of 100 μm between spots and spot centroids. Each spot contains millions of capture probes that bind to mRNA, and each probe contains a unique barcode (barcode). Principle of space transcriptome gene expression chip   Sample type: OCT-embedded samples that can be used for tissue slicing (not available for samples such as bone tissue)   Experimental process   Experimental flow of space transcriptome sequencing   Analysis process 　　Space transcriptome sequencing analysis process Application directions a. Tumor heterogeneity; b. Histomorphology; c. Tissue developmental mechanisms; d. Response to intervention; e. Biomarker discovery; f. Cellular mapping. Product advantages a. Comprehensive understanding of disease complexity b. Discovery of new biomarkers and identification of new cell types and states c. Locating the spatial structure of cellular profiles d. Identify spatio-temporal gene expression patterns

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