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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. [2] [3]
For example, to investigate a biological process which is estimated to occur for an hour, a researcher might design an experiment where the process is triggered for five minutes, 15 minutes, 30 minutes, 45 minutes, one hour, and two hours in separate cell culture samples before harvesting the cells for RNA-seq analysis.
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 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 sequencing is a next-generation sequencing technology; as such it requires only a small amount of RNA and no previous knowledge of the genome. [3] It allows for both qualitative and quantitative analysis of RNA transcripts, the former allowing discovery of new transcripts and the latter a measure of relative quantities for transcripts in a ...
Rail-RNA Scalable analysis of RNA-seq splicing and coverage. RAP RNA-Seq Analysis Pipeline, a new cloud-based NGS web application. RSEQtools "RSEQtools consists of a set of modules that perform common tasks such as calculating gene expression values, generating signal tracks of mapped reads, and segmenting that signal into actively transcribed ...
SHAPE has been used to analyze diverse RNA structures, including that of an entire HIV-1 genome. [25] The best approach is to use a combination of chemical probing reagents and experimental data. [26] In SHAPE-Seq SHAPE is extended by bar-code based multiplexing combined with RNA-Seq and can be performed in a high-throughput fashion. [27]
RACE can provide the sequence of an RNA transcript from a small known sequence within the transcript to the 5' end (5' RACE-PCR) or 3' end (3' RACE-PCR) of the RNA. This technique is sometimes called one-sided PCR or anchored PCR. The first step in RACE is to use reverse transcription to produce a cDNA copy of a region of the RNA transcript. In ...