fish(flourescent in-situ hybridization)
TRANSCRIPT
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FISH FLOURESCENT IN-SITU HYBRIDIZATION
NARENDRA YADAV PAPER 3 UNIT 2
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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.
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Protocol Outline
Preparation of the fluorescent probes
Denaturation of the probe and the target
Hybridization of the probe and the target
Detection
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DENATURE OF DNA
DENATURE OF PROBE
LABELLING WITH PROBE
HYBRIZATION OF DNA-PROBE
DETECTION
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Flourescent labled organism
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MicroRNA(green) and mRNA(red) visualization in differentiating C1C12 cells
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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 tO1000bp.
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PROBES
Stellaris(R) RNA FISH probes.
Fiber FISH.
Q-FISH.
Flow-FISH.
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APPLICATION
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.
Also used to detect circulating tumor cells, and tissues samples.
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Continue…
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.
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BIBLIOGRAPHY
GOOGLE-IMAGES
WIKIPEDIA
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THANK YOU