Dna Fish Protocol
Dna Fish Protocol - Related Videos
09:15
DNA-based Fish Species Identification Protocol
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Cited by 9 •
2010
This publication describes how to use the Agilent Fish Species Identification System to identify the species of a fish by extracting DNA and performing PCR and RFLP analysis.
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.
05:29
3D DNA FISH: A Technique to Locate a Specific Gene on a Chromosome
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2023
Circulating tumor cells (CTC) isolation and characterization from the blood of cancer patients is a type of non-invasive liquid biopsy and associated with relapse and disease progression. This video describes the technique of DNA fluorescence-in-situ-hybridization (DNA FISH) on CTCs captured on a medical wire, to study a specific gene of interest.
16:19
Fluorescence in situ hybridization (FISH) Protocol in Human Sperm
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Cited by 39 •
2009
This video-article describes, step by step, how to process a semen sample to achieve good-quality fluorescence in situ hybridization on human spermatozoa. Preparations obtained can be used for aneuploidy screening in the context of clinical diagnosis.
06:00
HCR-DNA FISH: A Fluorescence In Situ Hybridization Technique to Detect Viral DNA in Infected Cells
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2023
In this video, the cells infected with MCPyV virions were subjected to in situ hybridization chain reaction to detect MCPyV specific genomes. Further, the DNA-HCR technique was combined with FISH to visualize the MCPyV DNA in infected human skin cells.
FAQs
Specificity comes from the probe’s complementary base sequence, which can hybridize with the matching target after cellular DNA is denatured. Denaturation separates the DNA strands and exposes sequences for pairing, while fluorescent labeling makes the bound probe visible. This sequence-directed recognition allows investigators to distinguish selected genomic regions within individual fixed cells or chromosomes.
Denaturation makes target DNA accessible for probe hybridization, whereas stringent washing removes probes that are not sufficiently matched to their targets. These steps work together to improve the distinction between specific fluorescent signals and background fluorescence. Their effectiveness directly influences whether microscopy produces interpretable patterns for evaluating defined genomic sequences.
A signal’s position within a fixed cell or chromosome provides information about where a defined DNA sequence is located relative to the surrounding genomic structure. Examining these locations in individual cells supports analysis of spatial genome organization and cellular heterogeneity. The same approach can therefore connect sequence identity with its chromosomal or cellular context.
The workflow begins with fixed cells or chromosomes, followed by denaturation of cellular DNA. Fluorescently labeled probes are then allowed to hybridize with complementary target sequences. Stringent washes remove nonspecific or weakly associated probe, and fluorescence microscopy is used to visualize the remaining signals. Interpretation focuses on the presence and location of distinguishable fluorescent patterns.
Because the method identifies defined sequences in their cellular or chromosomal locations, signal positions can be used to map genes and examine chromosome structure. Altered signal arrangements may indicate chromosomal rearrangements, while comparisons of signal patterns can help reveal copy-number changes. These observations provide molecular cytogenetic evidence at the level of individual cells.
DNA FISH links specific genomic sequences to individual-cell patterns, making it useful when genomic organization or sequence status may vary among cells. In biology and medical research, it supports studies of development, disease mechanisms, chromosomal abnormalities, and cellular heterogeneity. Its microscopy-based readout preserves spatial information that is important for interpreting genome structure in context.