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    Fluorescence in Situ Hybridization: Presented By: Shreya Lodh Under The Guidance Of: Dr.T.R.Sharma Dr.R.K.Chahota

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    Asfoor gake1
    Fluorescence in situ hybridization (FISH) is a technique that uses fluorescent probes that bind only to specific DNA or RNA sequences. It allows researchers to identify and locate genetic material on chromosomes. FISH is useful for detecting chromosomal abnormalities, gene mapping, and analyzing structural chromosome changes. The procedure involves denaturing probes and chromosomes, hybridizing the probes to target sequences, and detecting fluorescence under a microscope. FISH has applications in diagnosing genetic conditions and comparing genomes between species.

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    Fluorescence in Situ Hybridization: Presented By: Shreya Lodh Under The Guidance Of: Dr.T.R.Sharma Dr.R.K.Chahota

    Uploaded by

    Asfoor gake1
    37 views20 pages
    Fluorescence in situ hybridization (FISH) is a technique that uses fluorescent probes that bind only to specific DNA or RNA sequences. It allows researchers to identify and locate genetic material on chromosomes. FISH is useful for detecting chromosomal abnormalities, gene mapping, and analyzing structural chromosome changes. The procedure involves denaturing probes and chromosomes, hybridizing the probes to target sequences, and detecting fluorescence under a microscope. FISH has applications in diagnosing genetic conditions and comparing genomes between species.

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    Fish method

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    Fluorescence in situ hybridization (FISH) is a technique that uses fluorescent probes that bind only to specific DNA or RNA sequences. It allows researchers to identify and locate genetic material on chromosomes. FISH is useful for detecting chromosomal abnormalities, gene mapping, and analyzing structural chromosome changes. The procedure involves denaturing probes and chromosomes, hybridizing the probes to target sequences, and detecting fluorescence under a microscope. FISH has applications in diagnosing genetic conditions and comparing genomes between species.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    37 views20 pages

    Fluorescence in Situ Hybridization: Presented By: Shreya Lodh Under The Guidance Of: Dr.T.R.Sharma Dr.R.K.Chahota

    Uploaded by

    Asfoor gake1
    Fluorescence in situ hybridization (FISH) is a technique that uses fluorescent probes that bind only to specific DNA or RNA sequences. It allows researchers to identify and locate genetic material on chromosomes. FISH is useful for detecting chromosomal abnormalities, gene mapping, and analyzing structural chromosome changes. The procedure involves denaturing probes and chromosomes, hybridizing the probes to target sequences, and detecting fluorescence under a microscope. FISH has applications in diagnosing genetic conditions and comparing genomes between species.

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    Fluorescence in situ

    Hybridization
    PRESENTED BY : SHREYA LODH
    UNDER THE GUIDANCE OF: DR.T.R.SHARMA
    DR.R.K.CHAHOTA
    INTRODUCTION
     FISH is a cytogenetic technique that uses
    fluorescent probes that bind to only those parts
    of the chormosomes with a high degree of
    sequences complementarity.

     Fluorescent probe-DNA complex can be studied


    under fluroscent microscope.

     Developed by the biomedical researchers in


    early’ 1980s.
    Characteristics of FISH
    FISH - a process which vividly paints
    chromosomes or portions of chromosomes
    with fluorescent molecules.

    identifies the presence and location of a


    region of DNA or RNA within morphologically
    preserved chromosome preparations, fixed
    cells or tissue sections
    (FISH) contd.
    Identifies chromosomal abnormalities

    Aids in gene mapping, toxicological


    studies, analysis of chromosome
    structural aberrations, and ploidy
    determination

    Centromere regions stained brighter -


    means they are rich in A-T bonds
    Advantage of (FISH)

    less labor-intensive method for


    confirming the presence of a
    DNA segment within an entire
    genome than other
    conventional methods like
    Southern blotting
    FISH: Limitations
    Probe design requires knowledge of
    specific chromosomal abnormalities to be
    studied
    Cutoff signals may be different among
    laboratories
    Processing errors, imperfect hybridization,
    non-specific binding, photobleaching,
    interobserver variability, and false positive
    and negative results are possible
    FISH Procedure
    Denature the chromosomes
    Denature the probe
    Hybridization
    Fluorescence staining
    Detection or store in the dark
    Probes
     Complementary sequences of target
    nucleic acids

     Designed against the sequence of interest.

     Probes are tagged with fluorescent dyes


    like biotin, fluorescein, Digoxigenin

     Size ranges from 20-40 bp to 1000bp.

     Stellaris(R) RNA FISH probes.


    FISH Procedure
    DENATURE OF
    PROBE

    DETECTION

    DENATURE
    OF DNA
    Interpretation of FISH

    Each fluorescently labeled probe that hybridizes


    to a cell nucleus in the tissue of interest will
    appear as a distinct fluorescent dot
     Diploid nuclei will have two dots
     If there is duplication in the region of
    interest, the gain will result in more than
    two dots
     If there is a loss in the region of interest,
    one or zero dot will result
    FISH Uses
    APPLICATIONS
    Also used in germ cell or prenatal diagnosis of
    conditions such as aneuploidies.
    Use to detect and localize the presence and absence of
    the specific DNA sequences on chormosomes.
    FISH often used for finding specific features in DNA
    for use in genetic counselling, medicine, and species
    identification.
    FISH can also be used to detect and localize specific
    RNA target (mRNA, IncRNA, miRNA) in cells.
    Diseases that are diagnosed using FISH include
    Angelman syndrome, 22q13 deletion syndrome,, acute
    lymphoblastic leukemia, Cri-du-chat, and Down
    syndrome.
    FISH can also be used to compare the genomes of two
    biological species, to deduce evolutionary relationships.
    Differences between genomes
    Major differences in the nature and amount of
    repetitive DNA

    • dpTa1 tandem repeat • 45S rDNA


    FISH and Telomeres
    Telomeric probes define the terminal
    boundaries of chromosomes (5’ and 3’ ends)
    Used in research of chromosomal
    rearrangements and deletions related to cell
    aging or other genetic abnormalities
    Special telomeric probes specific to individual
    chromosomes have been designed
    Probe is based on the TTAGGG repeat
    present on all human telomeres
    FISH and Telomeres
    FISH in Physical Mapping
    A physical map is primarily based on the locations of
    landmarks along a DNA molecule and units of distance are
    expressed in base pairs.

    Low Resolution Physical Mapping

     Cytogenetic map

     In situ hybridization
    Procedure
    THANK YOU …..

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