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An advanced alternative for RNA Sequencing of Individual Cryosections described above, RNA tomography (tomo-seq) features better RNA quantification and spatial resolution. [10] It is also based on tissue cryosectioning with further RNA sequencing of individual sections, yielding genome-wide expression data and preserving spatial information. [10]
Specific cell populations can be identified in two ways. First, by matching the transcriptome to previous single-cell RNA-seq (scRNA-seq) profiles for each cell type. [1] Second, using spatial differential expression (SpatialDE), a pattern recognition software that can differentiate tissue types without scRNA-seq data. [11]
FISSEQ combines the spatial context of RNA-FISH and the global transcriptome profiling of RNA-seq. [1] FISSEQ preserves the tissue allowing single molecule in situ RNA localization. The foundation of the method is a novel nucleic acid sequencing library construction method that stably cross-links cDNA amplicons within biological samples. [2]
The earliest RNA-Seq work was published in 2006 with one hundred thousand transcripts sequenced using 454 technology. [40] This was sufficient coverage to quantify relative transcript abundance. RNA-Seq began to increase in popularity after 2008 when new Solexa/Illumina technologies allowed one billion transcript sequences to be recorded.
RNA-Seq (named as an abbreviation of RNA sequencing) is a technique that uses next-generation sequencing to reveal the presence and quantity of RNA molecules in a biological sample, providing a snapshot of gene expression in the sample, also known as transcriptome. [2] [3]
RNA-seq measures the transcription of a specific gene by converting long RNAs into a library of cDNA fragments. The cDNA fragments are then sequenced using high-throughput sequencing technology and aligned to a reference genome or transcriptome which is then used to create an expression profile of the genes. [1]
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.
RNA sequencing (RNA-seq) RNA-seq is a high-throughput RNA sequencing technology that allows scientists to profile the entire RNA (transcriptome). Therefore, novel transcripts and gene expression level can be identified based on cDNA libraries. This method can be used for cancer diagnosis and treatment evaluation. [13]