Circular Rna Purification
Circular Rna Purification - Related Videos
14:26
Genome-wide Purification of Extrachromosomal Circular DNA from Eukaryotic Cells
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Cited by 59 •
2016
This paper presents a sensitive method called Circle-Seq for purifying extrachromosomal circular DNA (eccDNA). The method encompasses column purification, removal of remaining linear chromosomal DNA, rolling-circle amplification and high-throughput sequencing. Circle-Seq is applicable to genome-scale screening of eukaryotic eccDNA and studying genome instability and copy-number variation.
11:09
Chromatin Isolation by RNA Purification (ChIRP)
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Cited by 380 •
2012
ChIRP is a novel and rapid technique to map genomic binding sites of long noncoding RNAs (lncRNAs). The method takes advantage of the specificity of anti-sense tiling oligonucleotides to allow the enumeration of lncRNA-bound genomic sites.
02:30
Isolation of Small Regulatory RNA–Bound Bacterial Target RNA Using Affinity Purification
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2025
Source: Mercier, N., et. al., MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria. J. Vis. Exp. (2021)This video demonstrates the isolation of MS2-tagged small regulatory RNA (sRNA) bound to its bacterial target RNA using affinity purification with a maltose-binding protein (MBP)–MS2 coat protein fusion immobilized on amylose resin.
11:37
Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism
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Cited by 5 •
2017
This article provides a detailed procedure on the solid-phase synthesis, purification, and characterization of dodecamers of RNA modified at the C2'-O-position. UV-vis and circular dichroism photometric analyses are used to quantify and characterize structural aspects, i.e., single-strands or double-strands.
08:25
Identification of Circular RNAs using RNA Sequencing
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Cited by 30 •
2019
Circular RNAs (circRNAs) are non-coding RNAs that may have roles in transcriptional regulation and mediating interactions between proteins. Following assessment of different parameters for construction of circRNA sequencing libraries, a protocol was compiled utilizing stranded total RNA library preparation with RNase R pre-treatment and is presented here.
FAQs
Exonucleases are enzymes that digest linear RNA molecules from their exposed ends. Circular RNA lacks free 5′ and 3′ ends, so it shows greater resistance to this treatment than linear RNA. Applying exonuclease digestion can therefore reduce linear RNA in a sample and enrich the relative abundance of circular molecules before additional cleanup or separation steps.
Enzymatic treatment can reduce unwanted linear RNA, but it does not by itself remove every contaminant. Size-based separation and chromatography provide additional ways to improve sample purity by separating RNA from proteins, DNA, and other cellular components. Combining these approaches produces cleaner circular RNA for downstream structural, molecular, sequencing, or translational studies.
A useful preparation must address more than linear RNA. Proteins, DNA, and other cellular components can remain associated with the target material and interfere with downstream assays or interpretation. Purification workflows therefore use complementary treatments and separation methods to reduce these contaminants, helping ensure that observed results reflect circular RNA rather than impurities.
The lack of free 5′ and 3′ ends is the central biochemical feature used during enrichment. It makes circular molecules more resistant to exonucleases that act on linear RNA, creating a functional difference between the target and much of the background RNA. This distinction allows purification to begin with selective enzymatic removal before physical separation or chromatography.
A typical workflow first applies enzymatic treatment to exploit the target’s resistance to exonuclease digestion, then uses one or more cleanup methods. Size-based separation or chromatography can improve removal of proteins, DNA, residual RNA, and other cellular material. The resulting preparation is intended to provide cleaner circular RNA for later biochemical and molecular analyses.
Purified material is valuable when experiments require clearer analysis of circular RNA structure, stability, translation, or gene regulation. It also supports sequencing and molecular assays in which linear RNA or cellular contaminants could complicate interpretation. The same purification goal is relevant to research on RNA-based biomarkers and to the development of circular RNA therapeutics.
A cleaner preparation can support investigations of how circular RNA behaves as a molecule, including studies of its structure and stability, as well as its potential translation and regulatory roles. Purified samples also provide material for sequencing and other molecular assays. These applications connect biochemical isolation with broader research on biomarkers and therapeutic development.