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Single-cell sequencing examines the nucleic acid sequence information from individual cells with optimized next-generation sequencing technologies, providing a higher resolution of cellular differences and a better understanding of the function of an individual cell in the context of its microenvironment. [1]
This single cell shows the process of the central dogma of molecular biology, which are all steps researchers are interested to quantify (DNA, RNA, and Protein).. In cell biology, single-cell analysis and subcellular analysis [1] refer to the study of genomics, transcriptomics, proteomics, metabolomics, and cell–cell interactions at the level of an individual cell, as opposed to more ...
Single-cell omics technologies has extended beyond the transcriptome to profile diverse physical-chemical properties at single-cell resolution, including whole genomes/exomes, DNA methylation, chromatin accessibility, histone modifications, epitranscriptome (e.g., mRNAs, microRNAs, tRNAs, lncRNAs), proteome, phosphoproteome, metabolome, and more.
Detecting differences in gene expression level between two populations is used both single-cell and bulk transcriptomic data. Specialised methods have been designed for single-cell data that considers single cell features such as technical dropouts and shape of the distribution e.g. Bimodal vs. unimodal. [23]
TCR sequencing can be performed in on pooled cell populations (“bulk sequencing”) or single cells (“single cell sequencing”). [4] Bulk sequencing is useful to explore entire TCR repertoires - all the TCRs within an individual or a sample - and to generate comparisons between repertoires of different individuals. [4] This method can ...
Single-cell RNA sequencing (scRNA-Seq) provides the expression profiles of individual cells. Although it is not possible to obtain complete information on every RNA expressed by each cell, due to the small amount of material available, patterns of gene expression can be identified through gene clustering analyses .
The DNA sequencing is done on a chip that contains many ZMWs. Inside each ZMW, a single active DNA polymerase with a single molecule of single stranded DNA template is immobilized to the bottom through which light can penetrate and create a visualization chamber that allows monitoring of the activity of the DNA polymerase at a single molecule level.
Many technologies exist for sequencing genomes, but they are designed to use DNA from a population of cells rather than a single cell. The primary challenge for single-cell genome sequencing is to make multiple copies of (amplify) the DNA so that there is enough material available for the sequencer, a process called whole genome amplification ...