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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]
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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 ...
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. [24]
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.
The FAST4 format was invented as a derivative of the FASTQ format where each of the 4 bases (A,C,G,T) had separate probabilities stored. It was part of the Swift basecaller, an open source package for primary data analysis on next-gen sequence data "from images to basecalls". The FAST5 format was invented as an extension of the FAST4 format.
TCR-Seq (T-cell Receptor Sequencing) is a method used to identify and track specific T cells and their clones. [1] TCR-Seq utilizes the unique nature of a T-cell receptor (TCR) as a ready-made molecular barcode. [1] This technology can apply to both single cell sequencing technologies and high throughput screens [1]
Phred then uses these parameters to look up a corresponding quality score in huge lookup tables. These lookup tables were generated from sequence traces where the correct sequence was known, and are hard coded in Phred; different lookup tables are used for different sequencing chemistries and machines.