How does Ribo Zero work?
Similarly, the Ribo-Zero kit relies on biotinylated rRNA capture probes, which, following hybridization to the target rRNA molecules, are captured by magnetic beads. In contrast to MICROBExpress and RiboMinus, the RiboZero kit also targets 5S rRNA.
How do you get rid of ribosomal RNA?
Generally, one of two strategies is used to remove rRNA from total RNA, enrichment of mature polyadenylated (poly(A)) mRNA or targeted depletion of rRNA. The former is based on the use of oligo(dT) primers during reverse transcription of RNA into cDNA.
What is Ribo depletion?
Ribosomal depletion is a critical method in transcriptomics that allows for efficient detection of functionally relevant coding as well as non-coding transcripts through removal of highly abundant rRNA species.
How does rRNA depletion work?
The most common method of rRNA depletion is poly-A selection, which relies on the use of oligo (dT) primers attached to a solid support (e.g. magnetic beads) to isolate protein-coding polyadenylated RNA transcripts.
What is poly A enrichment?
Poly(A) enrichment is a very cost-efficient and fast pre-processing method that allows selection of mainly protein-coding mRNA. It can be used for all species that possess poly(A)-tailed RNA to remove undesired rRNA and concentrate sequencing reads on mRNA.
What is poly a capture?
The most popular RNA library used for RNA sequencing is the poly(A) captured RNA library. This library captures RNA based on the presence of poly(A) tails at the 3′ end. Another type of RNA library for RNA sequencing is the total RNA library which differs from the poly(A) library by capture method and price.
What is poly a selection?
The Poly(A) RNA Selection Module enables the rapid and highly specific enrichment of polyadenylated RNAs from total RNA samples.
Why do we need to get rid of ribosomal RNA for sequencing in an RNA seq experiment?
Ribosomal RNA (rRNA) is the most highly abundant component of total RNA isolated from animal or human cells and tissues, comprising the majority (>80% to 90%) of the molecules in a total RNA sample7. To allow efficient transcript/gene detection, highly abundant rRNAs must be removed from total RNA before sequencing.
How is RNA sequencing done?
RNA-seq involves conversion of a sample of RNA to a cDNA library, which is then sequenced and mapped against a reference genome. In addition to the ability to measure the level of gene expression, it provides further information on alternative splicing and non-coding RNA (such as microRNA) (Chaussabel et al., 2010).
What is poly-a selection?