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Single-cell RNA sequencing in immunology

Single-cell RNA sequencing (scRNA-seq) has revolutionized the study of the immune system and now has a wide range of applications in immunology. This technology spans the whole genome and provides an unbiased gene expression profile of individual cells. Bulk vs single-cell RNA-seq Traditional bulk RNA-seq is often performed on well-identified groups of cells thought to be homogeneous. However, quantification of molecular changes is made by estimating the mean value from millions of cells and averaging the signal of individual cells, thus ignoring cell-to-cell heterogeneity, which is a hallmark of adaptive immune cell subsets such as B and T lymphocytes. The need to identify new and discrete immune cell populations and to understand molecular changes that occur at the single cell level …

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Improving DNA amplification for single-cell genomics

Deep sequencing of genomes (Whole Genome Sequencing, WGS) is important not only to improve our knowledge in life sciences and evolutionary biology but also to make clinical progresses. The analysis of the genome and its variations at the cell level have major applications: analysis of mutation rates in somatic cells, including copy-number variations (CNVs)  and single-nucleotide variations (SNVs), evolution of cancer, recombination in germ cells, preimplantation genetic analysis for embryos or analysis of microbial populations (mini-metagenomics).   The single-cell DNA sequencing challenge Because of the low amount of DNA in a cell, single-cell whole genome sequencing requires whole genome amplification.  The 3 methods currently used are degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), multiple displacement amplification (MDA), and multiple annealing and looping-based …

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