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The single-cell RNA-Seq protocols vary in efficiency of RNA capture, which results in differences in the number of transcripts generated from each single cell. Single-cell libraries are usually sequenced to a depth of 1,000,000 reads because a large majority of genes are detected with 500,000 reads. [ 104 ]
RNA Seq Experiment. The single-cell RNA-seq technique converts a population of RNAs to a library of cDNA fragments. These fragments are sequenced by high-throughput next generation sequencing techniques and the reads are mapped back to the reference genome, providing a count of the number of reads associated with each gene. [13]
RNA-Seq can also be used to determine exon/intron boundaries and verify or amend previously annotated 5' and 3' gene boundaries. Recent advances in RNA-Seq include single cell sequencing, bulk RNA sequencing, [6] 3' mRNA-sequencing, in situ sequencing of fixed tissue, and native RNA molecule sequencing with single-molecule real-time sequencing. [7]
Some tools available to bulk RNA-Seq are also applied to single cell analysis, however to face the specificity of this technique new algorithms were developed. CEL-Seq [114] single-cell RNA-Seq by multiplexed linear amplification. Drop-Seq [115] Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets.
general repository for interaction datasets Protein-protein and other molecular interactions RNA-binding protein database: Protein-protein and other molecular interactions Database of Interacting Proteins: Univ. of California: Protein-protein and other molecular interactions IntAct [21] EMBL-EBI: open-source database for molecular interactions
Trajectory inference as implemented in Slingshot for (a) a simulated two-dimensional dataset and (b) a single-cell RNA-seq dataset of the olfactory epithelium.. Trajectory inference or pseudotemporal ordering is a computational technique used in single-cell transcriptomics to determine the pattern of a dynamic process experienced by cells and then arrange cells based on their progression ...
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