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A further consideration when sequencing large, branched cell types, such as neurons, comes from the removal of distal processes containing local pools of RNA during the single-cell isolation process. In these cells, scRNA-seq datasets only capture transcript in the central cell body, omitting transcripts from RNA pools localized to cellular ...
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. [ 105 ]
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.
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]
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).
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
Among the products of the program is the Azimuth reference datasets for single-cell RNA seq data [2] [3] and the ASCT+B Reporter, a visualization tool for anatomical structures, cell types and biomarkers. [4] [5] Millitomes are used to create uniformly sized tissue blocks that match the shape and size of organs from HuBMAP's 3D Reference Object ...
Open access across all immune system cells; expression data, differential expression, coregulated clusters, regulation: 267: 1059: Jan 2012 Genevestigator: Gene expression search engine based on manually curated, well annotated public and proprietary microarray and RNA-seq datasets: 3228: 232,855: October 2016 Gene Expression Omnibus - NCBI
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