circRNA Isolation Kit

Cat. No.

G4001

Unit

25 Reactions

Price

$550.00

Cat. No.

G4001

Name

circRNA Isolation Kit

Unit

25 Reactions

Description

abm’s circRNA Isolation Kit allows for the enrichment of a novel class of RNA called circular RNA from a total RNA sample. Poly(A) Polymerase, E. coli catalyzes the addition of adenosine residues to the 3’ termini of all linear RNAs – including structured RNAs. This results in RNAs exhibiting a long, 3’ unstructured overhang which is optimally bound by RNase R. RNase R can then illicit 3’ to 5’ exonuclease activity resulting in improved degradation of linear RNAs which contributes to higher purity circRNAs.

Product Component	Quantity
Poly(A) Polymerase, E. coli	25 µl
10X Poly(A) Polymerase, E. coli Reaction Buffer	1.0 ml
ATP (10 mM)	50 µl
RNA Purification Magnetic Beads	5 ml
RNase R	50 µl
10X RNase R Reaction Buffer	1.0 ml
RNaseOFF Ribonuclease Inhibitor	100 µl

Application

Enhanced circRNA isolation and enrichment.

Storage Condition

•Store all components except RNA Purification Magnetic Beads at -20°C
•Store RNA Purification Magnetic Beads at 4°C

Note

A magnetic separation rack is required (not included)

Material Citation

If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G4001

Datasheet

G4001 Datasheet

Search CoA here

MSDS

G4001 MSDS

How does the circRNA Isolation Kit method differ from other circRNA isolation methods?

circRNAs vary greatly by genesis and size from their linear RNA counterparts and thus cannot be purified from other RNAs by typical means of size or sequence. Instead they can be enriched by a series of treatments which can be applied in various orders. These include rRNA depletion, polyadenylation and/or RNase R treatment. Currently there are several common circRNA enrichment methods each with their own advantages and disadvantages, see our Learning Resources for a more detailed explanation. abm’s circRNA Isolation Kit utilizes a polyadenylation step followed by RNase R degradation.

Why does the circRNA Isolation Kit use magnetic purification beads instead of spin column purification?

circRNA are naturally present at low concentrations, comprising of only <0.1% of total RNA. As a result, achieving high recovery is crucial for downstream applications. This challenge is further compounded by the necessity of multiple enzymatic and clean up steps required in most circRNA isolation protocols which can lead to significant sample loss. abm’s RNA Purification Magnetic Beads (Cat. No. G971) offer a solution - they are paramagnetic beads that can reversibly bind nucleic acids and provide high recovery rates and reproducibility compared to traditional silica spin columns making them ideal for reliable circRNA isolation and purification.

	Can I use the circRNA Isolation Kit to directly isolate circRNA from my cells?
	No, you must first extract total RNA from your sample - abm recommends Column-Pure RNA Miniprep Kit (Cat. No. D518). The total RNA is then subjected to several enzymatic and clean up steps to isolate circRNA and remove the majority of other RNA classes.

	I'm interested in a couple components of the circRNA Isolation Kit. Are the kit components available separately?
	Yes, every component is available to purchase separately. Visit our main circRNA page to find all listed products.

	What type of magnetic bead is used to isolate the circRNA in the Isolation Kit?
	The kit includes paramagnetic beads coated with carboxyl groups that can reversibly bind to nucleic acids. The beads are intended for sample clean up in between enzymatic steps and provide high recovery rate and reproducibility.