To learn how to pre-process and analyze single-cell RNA-seq explore the following Google Colab notebooks that explain how to go from reads to results: Here we introduce some of the key concepts of single-cell RNA-seq technologies, with a focus on droplet based methods. Among such methods, single-cell RNA-seq (scRNA-seq) is having a profound impact on biology. The rapid development of single-cell genomics methods starting in 2009 has created unprecedented opportunity for highly resolved measurements of cellular states. ¶ *Division of Biology and Biological Engineering, California Institute of Technology ¶ Based on material taught in Caltech course Bi/BE/CS183 by Lior Pachter and Matt Thomson, with contributions from Sina Booeshaghi, Lambda Lu, Jialong Jiang, Eduardo Beltrame, Jase Gehring, Ingileif Hallgrímsdóttir and Valentine Svensson. *Division of Biology and Biological Engineering, California Institute of TechnologyĪn introduction to single-cell RNA-seq ¶ Written by Sina Booeshaghi* and Lior Pachter*. This service can be performed in conjunction with the 5′ Next GEM library prep or the VDJ immune profiling (B cell or T cell) library prep, but is not a stand alone service.Written by Sina Booeshaghi* and Lior Pachter*. It can also be used to multiplex more than one sample into a single library prep. This technology can also determine antigen specificity of single T cells with Feature Barcode peptide-MHC multimers to study the dynamic interactions between lymphocytes and antigens. Use this technique to measure both gene and cell surface protein expression in the same cell to identify protein isoforms, detect protein for low abundance transcripts, and further increase phenotypic specificity. Libraries are generated and sequenced and 10x Barcodes are used to associate individual reads back to the individual partitions. It does so by partitioning thousands of cells into nanoliter-scale Gel Beads-in-emulsion (GEMs), where all generated cDNA share a common 10x Barcode. A pool of ~750,000 barcodes are sampled separately to index each cell’s transcriptome and cell surface protein. This is accomplished by labeling cell surface proteins with antibodies conjugated to a Feature Barcode oligonucleotide, followed by direct capture of the Feature Barcode by the Gel Bead primer. The Chromium Single Cell Feature Barcode technology offers a comprehensive, scalable approach to detect cell surface proteins along with the gene expression and immune repertoire information from the same single cell. Plastic Surgery, Maxillofacial, & Oral Health.Translational Health Research Institute of Virginia.Institute of Law, Psychiatry & Public Policy.Child Health Research Center (Pediatrics).Thaler Center for AIDS & Human Retrovirus Research Center for Immunity, Inflammation & Regenerative Medicine.Center for Behavioral Health & Technology.Molecular Physiology & Biological Physics.Microbiology, Immunology, & Cancer Biology (MIC).
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |