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Single-cell DNA genome sequencing involves isolating a single cell, amplifying the whole genome or region of interest, constructing sequencing libraries, and then applying next-generation DNA sequencing (for example Illumina, Ion Torrent). Single-cell DNA sequencing has been widely applied in mammalian systems to study normal physiology and ...
scLVM [117] scLVM is a modelling framework for single-cell RNA-seq data that can be used to dissect the observed heterogeneity into different sources, thereby allowing for the correction of confounding sources of variation. scM&T-Seq Parallel single-cell sequencing.
Next generation sequencing machines have increased the rate of DNA sequencing substantially, as compared with the previous Sanger methods. DNA samples can be prepared automatically in as little as 90 mins, [5] while a human genome can be sequenced at 15 times coverage in a matter of days. [6]
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 .
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]
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.
CITE-Seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing) is a method for performing RNA sequencing along with gaining quantitative and qualitative information on surface proteins with available antibodies on a single cell level. [1] So far, the method has been demonstrated to work with only a few proteins per cell.
UMIs are particularly well-suited to single-cell RNA-Seq transcriptomics, where the amount of input RNA is restricted and extended amplification of the sample is required. [ 73 ] [ 74 ] [ 75 ] Once the transcript molecules have been prepared they can be sequenced in just one direction (single-end) or both directions (paired-end).