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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 ]
CITE-Seq. CITE-Seq (C ellular I ndexing of T ranscriptomes and E pitopes by Seq uencing) 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.
snRNA-seq, also known as single nucleus RNA sequencing, single nuclei RNA sequencing or sNuc-seq, is an RNA sequencing method for profiling gene expression in cells which are difficult to isolate, such as those from tissues that are archived or which are hard to be dissociated. It is an alternative to single cell RNA seq (scRNA-seq), as it ...
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]
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]
Geo-seq is a method that utilizes both laser capture microdissection and single-cell RNA sequencing procedures to determine the spatial distribution of the transcriptome in tissue areas approximately ten cells in size. [17] The workflow involves removal and cryosectioning of tissue followed by laser capture microdissection.