2-o-methyl Rna Detection
2-o-methyl Rna Detection - Related Videos
08:49
Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
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Cited by 7 •
2019
We present a detailed small RNA library reparation protocol with less bias than standard methods and an increased sensitivity for 2'-O-methyl RNAs. This protocol can be followed using homemade reagents to save cost or using kits for convenience.
10:19
High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum
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Cited by 15 •
2013
Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.
04:28
Multiplex Droplet Polymerase Chain Reaction to Detect DNA Methylation in Leukocytes
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2025
This video demonstrates the multiplex droplet polymerase chain reaction technique for the simultaneous quantification of methylated DNA regions in different white blood cell populations as an early biomarker for clinical diagnosis and prognosis.
09:12
DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
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2019
Here we present a protocol for DNAzyme-dependent cleavage of RNA. This enables fast and site-dependent analysis of RNA 2’-O-methylation. This approach can be used for the preliminary or major assessment of snoRNA activity.
10:34
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture
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Cited by 19 •
2016
An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.
FAQs
2′-O-methyl RNA detection relies on two distinct signatures of the modified ribose. A 2′-O-methyl group makes the nucleotide resistant to alkaline cleavage, while it can also hinder reverse-transcriptase movement during complementary-strand synthesis. Researchers translate either difference into a position-specific pattern, allowing modified sites to be mapped against surrounding RNA sequence information.
Resistance to alkaline cleavage creates a positional contrast within an RNA molecule. Unmodified nucleotides can produce cleavage patterns under alkaline treatment, whereas 2′-O-methylated positions resist that reaction. Comparing the resulting fragments therefore identifies protected sites and helps determine where modification occurs, although the interpretation depends on accurately relating cleavage positions to the RNA sequence.
During reverse transcription, a 2′-O-methylated nucleotide can impede the enzyme copying the RNA template. The resulting termination or reduced progression creates a detectable signal near the modified position. Primer-extension measurements or sequencing readouts can record this interruption, turning an enzymatic obstacle into a map of modification sites within the analyzed transcript.
Cleavage-based assays identify sites through resistance to RNA fragmentation, whereas primer-extension assays detect where reverse transcription is impeded. Sequencing readouts convert these positional effects into sequence-linked data that can be analyzed across an RNA molecule. The approaches therefore share the same chemical or enzymatic basis but differ in how site information is recorded and compared.
A general workflow selects an RNA target, applies a cleavage- or reverse-transcription-based detection strategy, and records the resulting fragments, extension patterns, or sequencing signals. Researchers then align those signals with the RNA sequence to identify candidate modified nucleotides. This workflow can be adapted to characterize individual RNAs or compare modification patterns across samples.
These approaches can be applied to ribosomal RNA, transfer RNA, and messenger RNA. Examining these different RNA classes helps investigators compare modification patterns across molecules with distinct cellular roles. The resulting maps can support studies of RNA processing and provide a basis for asking whether modification changes accompany differences in gene regulation or other biological states.
In genetics, site-specific maps connect RNA modifications with RNA-processing pathways and gene regulation. Researchers can also examine patterns during development or in disease-related molecular changes, then compare where modifications occur across RNA types or biological conditions. The measurements provide molecular evidence for investigating how altered RNA modification patterns may relate to genetic and cellular phenotypes.