FISH ( fluorescent in situ hybridization)
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FISH (Fluorescence in-situ Hybridization) is a technique that uses fluorescent probes to detect and localize specific DNA sequences on chromosomes. It can identify chromosomal abnormalities, aid in gene mapping, and diagnose diseases. The FISH procedure involves denaturing chromosomes and probes, hybridizing them, fluorescence staining, and examination under a microscope. FISH has advantages like being less labor-intensive than other methods and can detect submicroscopic deletions. It is used for research, prenatal diagnosis of conditions like aneuploidies, and diagnosing diseases such as Down syndrome.
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Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes to identify specific sequences on chromosomes. FISH allows researchers to detect the presence, absence, and location of gene sequences without having to culture cells. The process involves labeling probes with fluorescent dyes, applying the probes to intact cells on a slide, and visualizing where the probes bind to complementary DNA sequences under a microscope. FISH has applications in gene mapping, identifying chromosomal abnormalities, and analyzing structural chromosome changes.
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This presentation provides an insight into the fundamentals of in situ hybridization (ISH), especially fluorescence in situ hybridization. It is ideal for classroom lecture.
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Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes to identify specific sequences on chromosomes. FISH allows researchers to detect locations of genes on chromosomes, identify chromosomal abnormalities, and determine copy numbers of genes. The FISH process involves preparing fluorescent probes with sequences complementary to the target sequence, hybridizing the probes to denatured chromosomes, and visualizing the fluorescent signals to identify the probe locations on chromosomes. FISH has various applications in gene mapping, diagnosing genetic disorders, and studying microbial communities in environmental samples.
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Cytogenetics involves studying chromosomes and their relationship to cell behavior. Fluorescent in situ hybridization (FISH) is a powerful molecular cytogenetics technique that uses fluorescent probes to hybridize to specific DNA or RNA sequences, allowing localization of targets within cells. FISH can be used to detect genetic abnormalities, gene expression, and is widely used in medical applications such as diagnosing various cancers and genetic disorders.
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This document provides an overview of fluorescence in situ hybridization (FISH), a molecular cytogenetic technique used to detect and localize the presence or absence of specific DNA sequences on chromosomes. It discusses the history, basic principles, and procedures of FISH, including probe preparation and labeling, target denaturation and hybridization between probe and target, and fluorescence microscopy to detect signals. The document also outlines the major applications of FISH in research and clinical diagnosis, as well as its advantages in providing rapid analysis of cells and its limitations in only detecting predefined abnormalities with current probes.
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Fluorescence in situ hybridization (FISH) is a process that uses fluorescent probes that bind to specific parts of chromosomes to identify chromosomal abnormalities. FISH can identify the presence and location of specific DNA or RNA sequences on chromosomes within cells and tissues. It allows researchers to view specific chromosome segments or entire chromosomes with fluorescence microscopy. FISH is useful for applications like gene mapping, toxicology studies, detecting chromosomal structural abnormalities, and determining ploidy.
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Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes to identify specific sequences on chromosomes. FISH allows researchers to detect the presence, absence, and location of gene sequences without having to culture cells. The process involves labeling probes with fluorescent dyes, applying the probes to intact cells on a slide, and visualizing where the probes bind to complementary DNA sequences under a microscope. FISH has applications in gene mapping, identifying chromosomal abnormalities, and analyzing structural chromosome changes.
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Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes to identify specific sequences on chromosomes. FISH allows researchers to detect locations of genes on chromosomes, identify chromosomal abnormalities, and determine copy numbers of genes. The FISH process involves preparing fluorescent probes with sequences complementary to the target sequence, hybridizing the probes to denatured chromosomes, and visualizing the fluorescent signals to identify the probe locations on chromosomes. FISH has various applications in gene mapping, diagnosing genetic disorders, and studying microbial communities in environmental samples.
An introduction to fluorescent in situ hybridization.
Cytogenetics involves studying chromosomes and their relationship to cell behavior. Fluorescent in situ hybridization (FISH) is a powerful molecular cytogenetics technique that uses fluorescent probes to hybridize to specific DNA or RNA sequences, allowing localization of targets within cells. FISH can be used to detect genetic abnormalities, gene expression, and is widely used in medical applications such as diagnosing various cancers and genetic disorders.
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Fluorescent in-situ hybridization (FISH) is a technique that uses fluorescent probes that bind to only parts of the genome to detect and localize specific DNA sequences on chromosomes. It involves denaturing and hybridizing a fluorescent probe to a complementary DNA or RNA target sequence on preserved tissue. The bound probe can then be visualized with a fluorescent microscope to identify specific gene mutations, abnormalities, or other chromosome changes to aid in cancer diagnosis, prenatal testing, and other clinical applications.
Fluorescent in situ hybridization (FISH)
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
Fish
This document provides an overview of fluorescence in situ hybridization (FISH). It discusses:
1. FISH is a cytogenetic technique that uses fluorescent probes to bind specifically to chromosomal regions with complementary DNA sequences, which can then be viewed under a fluorescent microscope.
2. FISH can be used to identify chromosomal abnormalities, assist with gene mapping, and determine ploidy. It has advantages over other techniques like being less labor intensive.
3. Common applications of FISH include detecting aneuploidies for prenatal diagnosis, locating specific DNA sequences on chromosomes, and identifying features in DNA for uses like genetic counseling.
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FISH is a technique that uses fluorescent probes that bind to complementary DNA sequences on chromosomes to detect chromosomal abnormalities. It allows detection of structural changes like deletions and duplications not visible under a microscope. FISH gained recognition supporting genome mapping efforts. It involves labeling probes, denaturing probe and target DNA, and allowing hybridization. Results are then viewed under a fluorescence microscope. FISH has applications in detecting abnormalities, identifying marker chromosomes, and monitoring therapy. It provides advantages over traditional cytogenetics like rapid analysis of non-dividing cells.
FISH FISH
Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a complementary nucleotide sequence. It can be used to detect chromosomal abnormalities by identifying the presence or absence of specific DNA sequences on chromosomes. FISH utilizes fluorescent dyes that are visible under a fluorescence microscope to light up the probes, which allows researchers to see the chromosomal location of DNA sequences and determine abnormalities. It has applications in detecting various genetic diseases caused by chromosomal deletions or translocations.
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Molecular cytogenetics involves combining molecular biology techniques with cytogenetics. This document discusses several molecular cytogenetic techniques including karyotyping, FISH, CGH, and SKY. Karyotyping allows visualization of whole chromosome sets but cannot detect small structural abnormalities. FISH uses fluorescent probes to detect specific DNA sequences on chromosomes. CGH analyzes copy number variations by comparing test and reference DNA samples hybridized to probes. SKY simultaneously visualizes all chromosome pairs in different colors, making abnormalities easy to identify. These techniques are useful for detecting chromosomal abnormalities that can cause genetic disorders and economic losses if spread within animal populations. Preventing the dissemination of abnormalities through genetic testing is important.
Karyotyping
Karyotyping involves analyzing chromosomes to identify abnormalities. There are various banding techniques that stain chromosomes differently based on DNA composition, allowing identification. G-banding is commonly used, staining regions rich in adenine and thymine dark and regions rich in guanine and cytosine light. This reveals unique banding patterns for each chromosome. Fluorescent in situ hybridization (FISH) uses fluorescent probes to target specific DNA sequences, allowing analysis at the gene level.
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Comparitive genomic hybridisation
Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells
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In Situ Hybridization (ISH) is a technique that allows for precise localization of a specific segment of nucleic acid within a histologic section.
Fluorescence in situ hybridization (FISH)
FISH is a technique that uses fluorescent probes that bind to only those regions of DNA or RNA that have nucleotide sequences complementary to the probe's sequence. It allows researchers to visualize chromosomes and specific genetic loci. There are several types of FISH probes including whole chromosome painting probes, repetitive sequence probes, and locus-specific probes. FISH has many diagnostic applications such as detecting chromosomal abnormalities, genetic translocations, and deletions associated with conditions like Prader-Willi syndrome.
Cish
Chromogenic in situ hybridization (CISH) is a technique that combines in situ hybridization with immunohistochemistry to detect specific DNA sequences within tissue samples using brightfield microscopy. CISH uses nucleic acid probes labeled with enzymes that produce colored precipitates when exposed to chromogenic substrates, allowing visualization of gene amplification or rearrangements. CISH has advantages over fluorescence in situ hybridization (FISH) in being cheaper, more stable, and easier to use, though it is less sensitive than FISH. CISH is commonly used to assess HER2 gene amplification in breast cancer and detect genetic fusions in cancers.
Comparative genomic hybridization
Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that allows detection of copy number variations between a test and reference DNA sample without cell culturing. CGH involves labeling and hybridizing test and reference DNA to normal metaphase chromosomes before visualizing differences in fluorescence to identify regions of gains or losses. While CGH was originally used for cancer research, it can also detect chromosomal abnormalities associated with genetic disorders and has improved resolution over traditional cytogenetic methods. The main limitations of CGH are its inability to detect structural aberrations without copy number changes and resolutions above 5-10 megabases.
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Genome mapping
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Radioactive ISH
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Optimizing ISH Detection
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Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that compares the DNA of a test sample to a reference sample to detect copy number variations without cell culturing. It involves labelling the tumor and normal DNA with different fluorescent dyes, mixing them, and hybridizing them to normal chromosomes to detect losses or gains of genetic material in the tumor DNA through fluorescence ratios. While CGH can detect events over 10-20 Mb, array CGH uses genomic fragments as targets and can detect changes as small as 5-10 kb, making it a faster and more sensitive technique. Array CGH is commonly used in cancer research and diagnosis of genetic disorders.
fish
Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only parts of the chromosome with high sequence complementarity. It allows researchers to detect and localize the presence or absence of specific DNA sequences on chromosomes or specific genes. FISH utilizes fluorescent probes and fluorescence microscopy to identify the probes' location on chromosomes to detect chromosomal abnormalities. It provides results faster than conventional karyotyping and is used to diagnose various genetic conditions and cancers.
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Fluorescent in situ hybridization (FISH) is a powerful technique for detecting DNA or RNA sequences in cells by permitting labeled probes to interact with complementary sequences, allowing detection of specific sequences located on chromosomes. FISH has many applications in molecular biology and medical science, including gene mapping, diagnosis of chromosomal abnormalities, and studies of cellular structure and function. It can be used to detect aberrations in chromosome number as well as single locus targets through use of different types of probes.
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This document discusses fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH), which are molecular cytogenetic techniques used to localize DNA sequences on chromosomes. FISH uses fluorescent probes to detect specific DNA or RNA sequences on chromosomes. GISH uses total genomic DNA as a probe to detect specific chromosomes. Both techniques overcome limitations of conventional cytogenetics and have various applications, including gene mapping, analyzing structural abnormalities, and detecting aneuploidy. The document discusses the principles, methods, advantages and limitations of FISH and GISH.
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Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
Fish
This document provides an overview of fluorescence in situ hybridization (FISH). It discusses:
1. FISH is a cytogenetic technique that uses fluorescent probes to bind specifically to chromosomal regions with complementary DNA sequences, which can then be viewed under a fluorescent microscope.
2. FISH can be used to identify chromosomal abnormalities, assist with gene mapping, and determine ploidy. It has advantages over other techniques like being less labor intensive.
3. Common applications of FISH include detecting aneuploidies for prenatal diagnosis, locating specific DNA sequences on chromosomes, and identifying features in DNA for uses like genetic counseling.
fish- Fluorescence in situ hybridization
FISH is a technique that uses fluorescent probes that bind to complementary DNA sequences on chromosomes to detect chromosomal abnormalities. It allows detection of structural changes like deletions and duplications not visible under a microscope. FISH gained recognition supporting genome mapping efforts. It involves labeling probes, denaturing probe and target DNA, and allowing hybridization. Results are then viewed under a fluorescence microscope. FISH has applications in detecting abnormalities, identifying marker chromosomes, and monitoring therapy. It provides advantages over traditional cytogenetics like rapid analysis of non-dividing cells.
FISH FISH
Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a complementary nucleotide sequence. It can be used to detect chromosomal abnormalities by identifying the presence or absence of specific DNA sequences on chromosomes. FISH utilizes fluorescent dyes that are visible under a fluorescence microscope to light up the probes, which allows researchers to see the chromosomal location of DNA sequences and determine abnormalities. It has applications in detecting various genetic diseases caused by chromosomal deletions or translocations.
Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY
Molecular cytogenetics involves combining molecular biology techniques with cytogenetics. This document discusses several molecular cytogenetic techniques including karyotyping, FISH, CGH, and SKY. Karyotyping allows visualization of whole chromosome sets but cannot detect small structural abnormalities. FISH uses fluorescent probes to detect specific DNA sequences on chromosomes. CGH analyzes copy number variations by comparing test and reference DNA samples hybridized to probes. SKY simultaneously visualizes all chromosome pairs in different colors, making abnormalities easy to identify. These techniques are useful for detecting chromosomal abnormalities that can cause genetic disorders and economic losses if spread within animal populations. Preventing the dissemination of abnormalities through genetic testing is important.
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Karyotyping involves analyzing chromosomes to identify abnormalities. There are various banding techniques that stain chromosomes differently based on DNA composition, allowing identification. G-banding is commonly used, staining regions rich in adenine and thymine dark and regions rich in guanine and cytosine light. This reveals unique banding patterns for each chromosome. Fluorescent in situ hybridization (FISH) uses fluorescent probes to target specific DNA sequences, allowing analysis at the gene level.
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This document discusses various cytogenetic techniques used to study chromosomes, including their structure and abnormalities. It describes karyotyping to analyze all chromosomes for changes, as well as techniques like fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), spectral karyotyping (SKY), and multicolor FISH (M-FISH) that use fluorescent probes to detect DNA sequences on chromosomes. It also discusses banding techniques like G-banding and Q-banding that identify chromosomes based on dark and light bands corresponding to GC-rich and AT-rich regions. These techniques are used to diagnose chromosomal abnormalities and further cytogenetic research.
Comparitive genomic hybridisation
Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells
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In Situ Hybridization (ISH) is a technique that allows for precise localization of a specific segment of nucleic acid within a histologic section.
Fluorescence in situ hybridization (FISH)
FISH is a technique that uses fluorescent probes that bind to only those regions of DNA or RNA that have nucleotide sequences complementary to the probe's sequence. It allows researchers to visualize chromosomes and specific genetic loci. There are several types of FISH probes including whole chromosome painting probes, repetitive sequence probes, and locus-specific probes. FISH has many diagnostic applications such as detecting chromosomal abnormalities, genetic translocations, and deletions associated with conditions like Prader-Willi syndrome.
Cish
Chromogenic in situ hybridization (CISH) is a technique that combines in situ hybridization with immunohistochemistry to detect specific DNA sequences within tissue samples using brightfield microscopy. CISH uses nucleic acid probes labeled with enzymes that produce colored precipitates when exposed to chromogenic substrates, allowing visualization of gene amplification or rearrangements. CISH has advantages over fluorescence in situ hybridization (FISH) in being cheaper, more stable, and easier to use, though it is less sensitive than FISH. CISH is commonly used to assess HER2 gene amplification in breast cancer and detect genetic fusions in cancers.
Comparative genomic hybridization
Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that allows detection of copy number variations between a test and reference DNA sample without cell culturing. CGH involves labeling and hybridizing test and reference DNA to normal metaphase chromosomes before visualizing differences in fluorescence to identify regions of gains or losses. While CGH was originally used for cancer research, it can also detect chromosomal abnormalities associated with genetic disorders and has improved resolution over traditional cytogenetic methods. The main limitations of CGH are its inability to detect structural aberrations without copy number changes and resolutions above 5-10 megabases.
Chromosome analysis
Chromosome analysis evaluates the number and structure of a person's chromosomes to detect abnormalities. It can detect both numerical changes, such as an abnormal number of chromosomes, and structural changes, such as deletions or duplications. Techniques for chromosome analysis include fluorescent in-situ hybridization (FISH), comparative genomic hybridization (CGH), and array comparative genomic hybridization (array CGH). FISH uses fluorescent probes to detect specific chromosome segments or genes, while CGH and array CGH compare DNA samples to detect copy number variations between them. Chromosome analysis is used to diagnose conditions like Down syndrome, leukemia, and Prader-Willi syndrome.
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Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that compares the DNA of a test sample to a reference sample to detect copy number variations without cell culturing. It involves labelling the tumor and normal DNA with different fluorescent dyes, mixing them, and hybridizing them to normal chromosomes to detect losses or gains of genetic material in the tumor DNA through fluorescence ratios. While CGH can detect events over 10-20 Mb, array CGH uses genomic fragments as targets and can detect changes as small as 5-10 kb, making it a faster and more sensitive technique. Array CGH is commonly used in cancer research and diagnosis of genetic disorders.
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Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only parts of the chromosome with high sequence complementarity. It allows researchers to detect and localize the presence or absence of specific DNA sequences on chromosomes or specific genes. FISH utilizes fluorescent probes and fluorescence microscopy to identify the probes' location on chromosomes to detect chromosomal abnormalities. It provides results faster than conventional karyotyping and is used to diagnose various genetic conditions and cancers.
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FISH (fluorescence in situ hybridization) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a complementary nucleotide sequence. This allows researchers to see the chromosomal location of DNA sequences with a fluorescent microscope. FISH is useful for detecting chromosomal abnormalities, identifying genetic diseases caused by chromosomal deletions or translocations, and can detect certain genetic changes like the Philadelphia chromosome in chronic myeloid leukemia. It has many applications in research, clinical diagnostics, and determining genetic causes of developmental disabilities.
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In situ genetic technology was historically developed and mainly focused on detection purpose, allowing specific nucleic acid sequences to be visualized in morphologically preserved tissue sections. With the synergy of genetics and immunohistochemistry, in situ detection can correlate microscopic topological information with gene activity at the transcriptional or post-transcriptional levels in specific tissues. Furthermore, its resolution allows spatial distribution of nucleic acid products to be revealed in a heterogeneous cell population. The newest member to the franchise of in situ genetic technology is a direct-on-specimen enrichment methodology specifically for cell-free DNA liquid biopsy. Contrary to in situ detection, this in-well in situ innovation tackles the very first sample preparation step to reduce material loss, thereby improving overall sensitivity. Genomic nucleic acids purified from specimens have been proven to be time consuming and suffered from damages and losses; the evolution of in situ genetic technology offers a powerful tool for precision functional genomics, enabling cross-check between in vitro and in vivo findings. It further opens the door to ultimate genetic engineering in situ.
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Karyotyping is the process by which photographs of chromosomes are taken in order to determine the chromosome complement of an individual, including the number of chromosomes and any abnormalities.
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FISH (fluorescence in situ hybridization) is a molecular cytogenetic technique that uses fluorescent probes that bind to complementary DNA sequences on chromosomes. It can detect chromosomal abnormalities by localizing specific DNA sequences. Probes are labeled with fluorescent dyes and detected using fluorescence microscopy. FISH is useful for detecting various genetic abnormalities like translocations, deletions, and extra or missing chromosomes associated with diseases such as cancer, genetic syndromes, and neurological disorders.
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it will help to make easy the most difficult diagnosis, salivary gland tumor clearly in multiple aspects.
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FISH ( fluorescent in situ hybridization)
- 1. Presentation Topic: FISH (Fluorescence in-situ Hybridization) Subject: Advance In Medical Laboratory Muhammad Abbas BS MLT FSC MLT Rehman College of Allied Health sciences Hayatabad Peshawar
- 2. FISH
- 4. FISH (Fluorescent In Situ Hybridization) FISH - a process which vividly paints chromosomes or portions of chromosomes with fluorescent molecules FISH is a cytogenetic technique which can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
- 5. Fluorescence microscopy can be used to find out where the fluorescent probe bound to the chromosome. It uses fluorescent probes which bind only to those parts of the chromosome with which they show a high degree of sequence similarity.
- 6. Purpose of FISH Identifies chromosomal abnormalities Aids in gene mapping, toxicological studies, analysis of chromosome structural aberrations, and ploidy determination. Used to identify the presence and location of a region of DNA or RNA within morphologically preserved chromosome preparations, fixed cells or tissue sections
- 7. FISH Procedure Denature the chromosomes Denature the probe Hybridization Fluorescence staining Examine slides or store in the dark
- 9. FISH Uses Detection of high concentrations of base pairs Centromere regions stained brighter - means they are rich in A-T bonds Also used in germ cell or prenatal diagnosis of conditions such as aneuploidies (Aneuploidy is the presence of an abnormal number of chromosomes in a cell, for example a human cell having 45 or 47 chromosomes instead of the usual 46)
- 10. Uses Conti….. Used in research of chromosomal rearrangements and deletions related to cell aging or other genetic abnormalities Application in cytogenetics - can detect submicroscopic deletions and cryptic translocations of genes associated with unexplained mental retardation and miscarriages.
- 11. Uses Conti….. 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
- 12. Advantage less labor-intensive method for confirming the presence of a DNA segment within an entire genome than other conventional methods like Southern blotting
- 13. Probes Probes are the 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