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A typical human cell consists of about 2 x 3.3 billion base pairs of DNA and 600 million mRNA bases. Usually, a mix of millions of cells is used in sequencing the DNA or RNA using traditional methods like Sanger sequencing or next generation sequencing.
Single-cell transcriptomics uses sequencing techniques similar to single-cell genomics or direct detection using fluorescence in situ hybridization. The first step in quantifying the transcriptome is to convert RNA to cDNA using reverse transcriptase so that the contents of the cell can be sequenced using NGS methods as was done in genomics.
A list of more than 100 different single cell sequencing (omics) methods have been published. [1] The large majority of methods are paired with short-read sequencing technologies, although some of them are compatible with long read sequencing.
As a result of the aforementioned properties of single-cell transcriptomic data, batch correction methods developed for bulk sequencing data were observed to perform poorly. Consequently, researchers developed statistical methods to correct for batch effects that are robust to the properties of single-cell transcriptomic data to integrate data ...
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 .
Analysis of single-cell sequencing presents many challenges, such as determining the best way to normalize the data. [8] Due to a new level of complications that arise from sequencing of both proteins and transcripts at a single-cell level, the developers of CITE-Seq and their collaborators are maintaining several tools to help with data analysis.
Two methods for single-cell ATAC-seq [8]. ATAC-seq stands for Assay for Transposase-Accessible Chromatin with high throughput sequencing. [9] It is a technique used in molecular biology to identify accessible DNA regions, equivalent to DNase I hypersensitive sites. [9]
Within the past five years, the development of single-cell Hi-C has enabled the depiction of the entire 3D structural landscape of chromatins/chromosomes throughout the cell cycle, and many studies have discovered that these identified genomic domains remain unchanged in interphase, and are erased by silencing mechanisms when the cell enters ...
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