Dig Labeled Rna Probe
Dig Labeled Rna Probe - Related Videos
11:15
In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA
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Cited by 16 •
2015
Here a method to localize bacteria within paraffin-embedded tissues using DIG-labeled 16S rRNA-targeting DNA probes has been described. This protocol can be applied to study the role of bacteria in various diseases such as periodontitis, cancers, and inflammatory immune diseases.
03:31
Labeling DNA Probes
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2021
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions. Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
12:16
Robust 3D DNA FISH Using Directly Labeled Probes
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Cited by 49 •
2013
We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.
03:26
Probe Hybridization and Signal Amplification in RNA In Situ Hybridization: A Technique for Detecting Specific RNA Sequences in Tissue Sections
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2023
This video describes a sequential hybridization strategy that involves hybridizing mRNA probes to specific target mRNA. Following probe hybridization, signal amplification is performed to enhance resolution via reduction of signal-to-noise ratio during RNA-CISH of histological samples.
06:08
In Situ Hybridization Using Oligonucleotide Probes to Study RNA Expression in Rat Brain Sections
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2025
This video demonstrates the detection of serotonin receptor RNA synthesis using oligonucleotide probes in rat brain tissue affected by serotonin syndrome. The protocol includes tissue pretreatment, probe hybridization, and enzymatic detection to visualize and quantify serotonin receptor RNA expression under a microscope.
FAQs
During probe synthesis, digoxigenin is incorporated into the RNA while the probe sequence is generated for a specific target. This creates a detectable molecular tag without changing the central recognition step: complementary base sequences pair during hybridization. The sequence therefore determines which RNA or DNA molecule can be detected in the sample.
Hybridization temperature and salt concentration help control whether the probe remains paired with its complementary target. Because the pairing occurs under controlled conditions, these variables influence how selectively the probe recognizes matching sequences. In practice, adjusting them is important when the goal is to distinguish the intended transcript or DNA region from less closely matching material.
Washing is a separation step rather than a labeling step. It removes probe that did not remain bound after hybridization, reducing signal from unbound material. The remaining DIG label is then revealed through an anti-DIG antibody connected to an enzyme or fluorescent reporter, linking molecular recognition to an observable signal.
Compared with radioactive probes, DIG-labeled RNA probes provide a nonradioactive alternative while retaining sequence-specific detection. Their readout depends on antibody recognition of DIG and a linked enzyme or fluorescent reporter, rather than on a radioactive label. This makes the labeling strategy relevant when researchers need to visualize complementary molecules without radioactive probe detection.
A typical workflow starts by synthesizing the RNA probe with DIG, then exposing a biological sample to the probe under controlled hybridization conditions. Washing follows to remove unbound material, and anti-DIG detection supplies the readout through an enzyme or fluorescent reporter. This sequence connects probe preparation, target binding, background reduction, and signal generation in one assay.
In developmental biology, these probes are especially useful for locating transcripts within embryos, cells, or tissues. In situ hybridization can show where a complementary RNA target is present, while gene expression analysis uses the resulting signal to examine transcript distribution. The method therefore provides spatial information about transcript presence in particular biological samples and locations.