The technique of fluorescence in situ hybridization involves the use of a ________ that hybridizes to a ________.

Question

BlackTom AI · Accepted Answer

Fluorescence in situ hybridization (FISH) uses fluorescently labeled nucleic acid probes that bind to complementary DNA sequences on intact chromosomes fixed on a slide.

1) Relevant knowledge points and concepts
- "In situ" means in the original place: DNA remains in chromosome form on a microscope slide rather than separated by electrophoresis.
- FISH uses fluorescent labels (e.g., fluorophores) on probes so hybridized sites can be visualized by fluorescence microscopy.
- Different probe types (short oligonucleotides, cDNA, BAC clones) target specific loci, whole chromosomes, or repetitive sequences to detect gene location, copy number, or structural rearrangements.

2) Step-by-step reasoning process
- Prepare cells and make chromosome spreads on a slide (preserves chromosomes in situ).
- Denature chromosomal DNA so single-stranded target sequences are available.
- Apply a complementary DNA/RNA probe labeled with a fluorophore.
- Allow hybridization; the probe binds to its complementary sequence at the specific site on the intact chromosome.
- Wash off unbound probe and visualize fluorescence at the hybridized site using a fluorescence microscope.

3) Intermediate steps (method sequence)
1. Cell fixation and slide preparation.
2. DNA denaturation.
3. Addition of fluorescently labeled probe.
4. Hybridization incubation.
5. Washing to remove non-specific binding.
6. Fluorescence detection and imaging.

4) Why the correct answer is correct
The correct answer (d) matches FISH by specifying a fluorescent probe and a target that is a specific site on an intact chromosome. FISH’s defining features are the fluorescent label and the in situ detection of sequences on chromosomal preparations.

5) Why other options are incorrect
- (a) Radiolabeled probe to a band on a gel: Radiolabels and gel bands describe Southern blotting or gel hybridization, not FISH.
- (b) Radiolabeled probe to an intact chromosome: Using radiolabels on intact chromosomes is not standard FISH methodology; visualization by light/fluorescence microscopy requires fluorescent labels.
- (c) Fluorescent probe to a band on a gel: Fluorescent probes can be used on blots, but "in situ" implies intact chromosomes, not gel bands.

6) Conclusion
The correct choice is d) fluorescent probe, specific site on an intact chromosome, which matches the provided answer.

The technique of fluorescence in situ hybridization involves the use of a that hybridizes to a .单项选择题

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The technique of fluorescence in situ hybridization involves the use of a ________ that hybridizes to a ________.单项选择题