Rna Labeling
Rna Labeling - Related Videos
04:10
Helicase Activity Measurement of a Target Protein Using Biotin-Labeled RNA Duplexes
0 Views •
2025
In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.
11:00
Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
0 Views •
Cited by 105 •
2013
Total cellular RNA provides a poor template for studying short-term changes in RNA synthesis and decay as well as the kinetics of RNA processing. Here, we describe metabolic labeling of newly transcribed RNA with 4-thiouridine followed by thiol-specific biotinylation and purification of newly transcribed RNA allowing to overcome these limitations.
05:34
Denaturing Urea Polyacrylamide Gel Electrophoresis for RNA Analysis: A Technique to Separate Fluorescently Labeled Phosphorylated RNA Oligonucleotides from their Non-Phosphorylated Equivalents
0 Views •
2025
In this video, we demonstrate a denaturing polyacrylamide gel electrophoresis for the separation of phosphorylated RNA species from their non-phosphorylated counterparts. The RNA species are fluorescently labeled for laser detection on the gel.
09:59
Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation
0 Views •
Cited by 14 •
2018
This method can be used to measure RNA synthesis. 5-Bromouridine is added to cells and incorporated into synthesized RNA. RNA synthesis is measured by RNA extraction immediately after labelling, followed by 5-Bromouridine-targeted immunoprecipitation of labelled RNA and analysis by reverse transcription and quantitative polymerase chain reaction.
08:28
Metabolic Labeling
0 Views •
2023
Metabolic labeling is used to probe the biochemical transformations and modifications that occur in a cell. This is accomplished by using chemical analogs that mimic the structure of natural biomolecules. Cells utilize analogs in their endogenous biochemical processes, producing compounds that are labeled. The label allows for the incorporation of detection and affinity tags, which can then be used to elucidate metabolic pathways using other biochemical analytical techniques, such as SDS-PAGE...
FAQs
The labeling route determines when and how the tag becomes associated with RNA. Modified nucleotides can be incorporated during transcription, chemical reactions can add a label after synthesis, and complementary probes can bind a chosen RNA sequence. These alternatives distinguish labels introduced into newly produced molecules from labels detected through sequence recognition, helping match the approach to the biological question.
The tag controls the signal used for measurement. Fluorescent labels support detection based on emitted light, affinity tags enable purification of associated RNA, enzymatic labels generate a signal through enzyme activity, and sequencing-based approaches provide sequence information about labeled molecules. Selecting among these readouts determines whether the experiment emphasizes visualization, isolation, signal generation, or molecular identification.
Complementary probes provide sequence-directed recognition rather than relying only on a tag incorporated into the RNA. Because a probe binds a specific RNA sequence, this strategy can focus detection on a selected transcript or RNA region. That distinction matters when the biological aim is to examine one RNA species within a larger collection of cellular RNA molecules.
The same label does not by itself define the biological conclusion; the measured feature depends on experimental design. Signal abundance can support abundance studies, spatial detection can address localization, movement measurements can address RNA movement, and capture or sequencing can examine interactions or molecular identities. Thus, labeling connects a physical readout to a specific RNA behavior.
A practical workflow begins by identifying whether the goal is abundance, location, movement, interactions, processing, or stability. The researcher then selects incorporation during transcription, post-synthesis chemistry, or complementary-probe binding, followed by a compatible readout such as fluorescence, affinity purification, enzymatic signal generation, or sequencing. This sequence keeps the strategy aligned with the intended measurement.
In biology, RNA labeling can follow gene expression and RNA processing, assess stability, and examine subcellular localization or movement. It also supports studies of molecular interactions by enabling detection or purification of labeled RNA. These applications help researchers relate RNA behavior to cellular function rather than treating RNA abundance as the only relevant measurement.
By making RNA behavior measurable, labeling approaches can connect altered abundance, localization, processing, stability, or interactions with changes in cellular function. This is relevant to disease-mechanism research because it broadens analysis beyond whether an RNA is present. The resulting measurements can help characterize how RNA-related changes are associated with biological dysfunction.