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This volume provides a comprehensive overview for investigating biology at the level of individual cells. Chapters are organized into eight parts detailing a single-cell lab, single cell DNA-seq, RNA-seq, single cell proteomic and epigenetic, single cell multi-omics, single cell screening, and single cell live imaging. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Single Cell Methods: Sequencing and Proteomics aims to make each experiment easily reproducible in every lab.
List of contents
Setting Up a Single-Cell Genomic Laboratory.- Tissue Handling and Dissociation for Single-Cell RNA-seq.- Full-Length Single-Cell RNA-Sequencing with Smart-seq2.- CEL-Seq2 - Single Cell RNA Sequencing by Multiplexed Linear Amplification.- Single-Cell RNA-seq by Multiple Annealing and Tailing Based Quantitative Single-Cell RNA-seq (MATQ-seq).- Single-Cell RNA Sequencing with Drop-Seq.- Chromium 10X Single Cell 3' mRNA Sequencing of Tumor Infiltrating Lymphocytes.- Seq-Well: A Sample-Efficient, Portable Picowell Platform for Massively Parallel Single-Cell RNA-Sequencing.- Single-Cell Tagged Reverse Transcription (STRT-seq).- Single-Cell RNA-Sequencing of Peripheral Blood Mononuclear Cells with ddSEQ.- High Throughput Single Cell Real-Time Quantitative PCR Analysis.- Single-Cell Dosing and mRNA Sequencing of Suspension and Adherent Cells Using the Polaris(TM) System.- Targeted TCR Amplification from Single-Cell cDNA Libraries.- Sequencing the Genomes of Single Cells.- Studying DNA Methylation in Single Cell Format with scBS-seq.- Single Cell 5fC Sequencing.- ChIPmentation for Low-Input Profiling of In Vivo Protein-DNA Interactions.- Immuno-Phenotyping of Human Peripheral Blood Mononuclear Cells by Mass Cytometry.- Classification of the Immune Composition in the Tumor Infiltrate.- Combined Genome and Transcriptome (G&T) Sequencing of Single Cells.- Simultaneous Pofiling of mRNA Transcriptome and DNA Methylome from a Single Cell.- Simultaneous Targeted Detection of Proteins and RNAs in Single Cells.- CRISPR-Screening in Single Cells.- Single Cell Live Imaging.- Differential Expression Analysis in Single Cell Transcriptomics.- A Bioinformatic Toolkit for Single-Cell mRNA Analysis.
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