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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.
The word transcriptome is a portmanteau of the words transcript and genome.It appeared along with other neologisms formed using the suffixes -ome and -omics to denote all studies conducted on a genome-wide scale in the fields of life sciences and technology.
In the 1980s, low-throughput sequencing using the Sanger method was used to sequence random transcripts, producing expressed sequence tags (ESTs). [2] [14] [15] [16] The Sanger method of sequencing was predominant until the advent of high-throughput methods such as sequencing by synthesis (Solexa/Illumina).
Fluorescent in situ sequencing (FISSEQ), [49] like ISS padlock, is a method that uses reverse transcription, rolling-circle amplification, and sequencing by ligation techniques. [5] It allows spatial transcriptome analysis in fixed cells. [ 5 ]
This difference results in strong batch effects that may bias the findings of statistical methods applied across batches, particularly in the presence of confounding. [30] As a result of the aforementioned properties of single-cell transcriptomic data, batch correction methods developed for bulk sequencing data were observed to perform poorly.
G&T-seq (short for single cell genome and transcriptome sequencing) is a novel form of single cell sequencing technique allowing one to simultaneously obtain both transcriptomic and genomic data from single cells, allowing for direct comparison of gene expression data to its corresponding genomic data in the same cell...
De novo transcriptome assembly is often the preferred method to studying non-model organisms, since it is cheaper and easier than building a genome, and reference-based methods are not possible without an existing genome. The transcriptomes of these organisms can thus reveal novel proteins and their isoforms that are implicated in such unique ...
[1] [2] Deep sequencing refers to the general concept of aiming for high number of unique reads of each region of a sequence. [3] Physical coverage, the cumulative length of reads or read pairs expressed as a multiple of genome size. [4] Genomic coverage, the percentage of all base pairs or loci of the genome covered by sequencing.