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Fluorescence in Situ Hybridization Video Lecture | Crash Course for CSIR NET Life Sciences

Q: 3. What are the major applications of the FISH technique in biological research and medicine?

Ans. FISH has a wide range of applications including:1. Cytogenetics, for detecting chromosomal abnormalities in cancers or genetic disorders.2. Gene mapping, to locate genes on chromosomes.3. Detection of specific RNA molecules in cells, aiding in understanding gene expression.4. Assessment of microbial communities through fluorescent probes targeting specific DNA sequences in environmental samples.5. Clinical diagnostics, for identifying genetic diseases and prenatal screening.

Q: 4. What are the advantages of using the FISH technique compared to other genetic analysis methods?

Ans. The advantages of FISH include:1. High specificity and sensitivity in detecting specific nucleic acid sequences.2. The ability to visualize multiple targets simultaneously using different fluorescent probes.3. Capability to analyze preserved tissues and cells, maintaining the spatial context of the genetic information.4. Rapid results compared to traditional karyotyping methods which can be time-consuming.5. Less complex sample preparation and analysis compared to some molecular techniques.

Q: 5. What limitations should researchers be aware of when using FISH?

Ans. Some limitations of FISH include:1. The requirement for high-quality fixed samples, as poor fixation can lead to non-specific binding or loss of target sequences.2. Limited resolution compared to techniques like single-cell sequencing, making it difficult to detect small genomic rearrangements.3. The need for specialized equipment and expertise in fluorescence microscopy.4. Potential for probe design challenges, as the specificity and efficiency depend on the quality and binding characteristics of the probes used.5. It may not provide quantitative data on gene expression levels without additional techniques.

FAQs on Fluorescence in Situ Hybridization Video Lecture - Crash Course for CSIR NET Life Sciences

1. What is the principle behind the Fluorescence In Situ Hybridization (FISH) technique?

Ans. The principle of FISH involves the use of fluorescently labeled DNA or RNA probes that hybridize to specific complementary sequences in the target nucleic acids within fixed cells or tissue sections. When exposed to light of a specific wavelength, these probes emit fluorescence, allowing for the visualization and localization of specific genetic material, making it possible to analyze chromosomal structure, number, and arrangement.

2. What are the key steps involved in the FISH procedure?

Ans. The FISH procedure generally includes several key steps: 1. Sample preparation, where cells or tissue sections are fixed onto a slide. 2. Denaturation of the DNA or RNA to separate the strands. 3. Hybridization, where fluorescent probes are added to the sample and allowed to bind to their complementary sequences. 4. Washing to remove unbound probes. 5. Detection and visualization using fluorescence microscopy, allowing for the analysis of the hybridized probes.

3. What are the major applications of the FISH technique in biological research and medicine?

Ans. FISH has a wide range of applications including: 1. Cytogenetics, for detecting chromosomal abnormalities in cancers or genetic disorders. 2. Gene mapping, to locate genes on chromosomes. 3. Detection of specific RNA molecules in cells, aiding in understanding gene expression. 4. Assessment of microbial communities through fluorescent probes targeting specific DNA sequences in environmental samples. 5. Clinical diagnostics, for identifying genetic diseases and prenatal screening.

4. What are the advantages of using the FISH technique compared to other genetic analysis methods?

Ans. The advantages of FISH include: 1. High specificity and sensitivity in detecting specific nucleic acid sequences. 2. The ability to visualize multiple targets simultaneously using different fluorescent probes. 3. Capability to analyze preserved tissues and cells, maintaining the spatial context of the genetic information. 4. Rapid results compared to traditional karyotyping methods which can be time-consuming. 5. Less complex sample preparation and analysis compared to some molecular techniques.

5. What limitations should researchers be aware of when using FISH?

Ans. Some limitations of FISH include: 1. The requirement for high-quality fixed samples, as poor fixation can lead to non-specific binding or loss of target sequences. 2. Limited resolution compared to techniques like single-cell sequencing, making it difficult to detect small genomic rearrangements. 3. The need for specialized equipment and expertise in fluorescence microscopy. 4. Potential for probe design challenges, as the specificity and efficiency depend on the quality and binding characteristics of the probes used. 5. It may not provide quantitative data on gene expression levels without additional techniques.

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Sep 28, 2025 Last updated

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