Probe Hybridization
Probe Hybridization - Related Videos
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.
08:49
Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome
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Cited by 2 •
2015
This protocol describes a rapid and simplified in situ hybridization method ideal forparaformaldehyde-prefixed brain, thus reducing the need for prolonged complex steps while using fresh frozen tissues. The method is validated using the identification of the serotonin 5-HT2A receptor gene htr2a in rats.
10:01
Observation and Quantification of Telomere and Repetitive Sequences Using Fluorescence In Situ Hybridization (FISH) with PNA Probes in Caenorhabditis elegans
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Cited by 5 •
2016
We report a concise procedure of fluorescence in situ hybridization (FISH) in the gonad and embryos of Caenorhabditis elegans for observing and quantifying repetitive sequences. We successfully observed and quantified two different repetitive sequences, telomere repeats and template of alternative lengthening of telomeres (TALT).
08:00
Whole-Mount In Situ Hybridization
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2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
FAQs
Selectivity comes from the probe’s nucleic acid sequence matching the complementary bases in a target DNA or RNA sequence. Only targets with the appropriate complementary arrangement provide a basis for stable pairing under the chosen conditions. This sequence-directed recognition allows researchers to locate or identify particular nucleic acid regions within a larger biological sample.
These conditions regulate how selectively the probe remains paired with its target. Temperature and salt concentration influence the stability of probe-target binding, while washing stringency removes less stable associations. Changing these variables can therefore alter both binding specificity and signal strength, which is important when distinguishing a desired sequence from other nucleic acids.
The label provides the detectable signal that reveals where the probe has bound. After complementary pairing and washing, the observed signal indicates the presence or location of the matching DNA or RNA sequence. Its role connects an otherwise molecular-scale interaction to measurable results in assays used for sequence identification, localization, or measurement.
The same pairing principle can be applied to either type of nucleic acid, but the biological question changes with the target. Southern blotting supports analysis of DNA organization, whereas Northern blotting is used in the context of RNA and gene expression. This flexibility makes the technique relevant to both genome structure and cellular activity.
A typical workflow selects a probe complementary to the sequence of interest, applies a label for detection, allows pairing under controlled conditions, and then uses washing to regulate which associations remain. The resulting signal is interpreted as evidence of the target sequence. Temperature, salt concentration, and washing conditions must be controlled because they affect the outcome.
These formats use the same sequence-recognition principle for different analytical purposes. Southern blots examine DNA, Northern blots address RNA, fluorescence in situ hybridization visualizes targets in chromosomes or cells, and microarrays support analysis across many sequence targets. The selected format determines whether the main outcome concerns organization, expression, localization, or broader sequence measurement.
The technique is useful when researchers need to detect, locate, identify, or measure a specific nucleic acid sequence. Applications include studying gene organization and expression, detecting pathogens or genetic variants, and visualizing chromosome or cellular targets. Diagnostic assays extend these capabilities to identifying biologically important sequence signals in samples.