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3' mRNA-seq is a quantitative, genome-wide transcriptomic technique based on the barcoding of the 3' untranslated region (UTR) of mRNA molecules. Unlike standard bulk RNA-seq, where short sequencing reads are generated along the entire length of mRNA transcripts, only the 3' end of polyadenylated RNAs are sequenced in 3' mRNA-seq.
Schematic overview of the MERCURIUS BRB-seq workflow where up to 384 samples can be barcoded and multiplexed per kit.. Bulk RNA barcoding and sequencing (BRB-seq) is an ultra-high-throughput bulk 3' mRNA-seq technology that uses early-stage sample barcoding and unique molecular identifiers (UMIs) to allow the pooling of up to 384 samples in one tube early in the sequencing library preparation ...
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 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]
It is important to distinguish the 5' and 3' UTRs from other non-protein-coding RNA. Within the coding sequence of pre-mRNA, there can be found sections of RNA that will not be included in the protein product. These sections of RNA are called introns. The RNA that results from RNA splicing is a sequence of exons. The reason why introns are not ...
Sequencing technology platforms commonly used for RNA-Seq [77] [78] Platform Commercial release Typical read length Maximum throughput per run Single read accuracy RNA-Seq runs deposited in the NCBI SRA (Oct 2016) [79] 454 Life Sciences: 2005 700 bp 0.7 Gbp 99.9% 3548 Illumina: 2006 50–300 bp 900 Gbp 99.9% 362903 SOLiD: 2008 50 bp 320 Gbp 99. ...
(See Central dogma of molecular biology). mRNA structure, approximately to scale for a human mRNA, where the median length of 3′UTR is 700 nucleotides. In molecular genetics, the three prime untranslated region (3′-UTR) is the section of messenger RNA (mRNA) that immediately follows the translation termination codon.
MPSS allows mRNA transcripts to be identified through the generation of a 17–20 bp signature sequence adjacent to the 3'-end of the 3'-most site of the designated restriction enzyme (commonly Sau3A or DpnII). Each signature sequence is cloned onto one of a million microbeads.