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Following are the steps of southern blot...

Following are the steps of southern blot procedure.
I. Autoradiography
II. Hybridization with radioactive nucleic acid (probe)
III. Blotting
IV. DNA fragments are treated to make them single stranded
V. Electrophoresis
VI. Clearing of DNA by restriction endonuclease
VII. Isolation of DNA from sample
The correct sequence is-

A

1, 2, 3, 4, 5, 6, 7

B

7, 6, 5, 4, 3, 2, 1

C

1, 2, 6, 7, 3, 4, 5

D

7, 6, 1, 2, 5, 3, 4

Text Solution

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The correct Answer is:
To solve the question regarding the correct sequence of steps in the Southern blot procedure, we can break down the process into logical steps based on the information provided. Here’s the step-by-step solution: ### Step-by-Step Solution: 1. **Isolation of DNA from Sample (VII)**: The first step in the Southern blot procedure is to isolate the DNA from the biological sample. This involves extracting the DNA to be analyzed. 2. **Clearing of DNA by Restriction Endonuclease (VI)**: Once the DNA is isolated, it is treated with restriction enzymes to cut the DNA into smaller fragments. This step is crucial for the subsequent analysis. 3. **Electrophoresis (V)**: After the DNA fragments are generated, they are separated by size using gel electrophoresis. This technique allows for the visualization of the DNA fragments based on their size. 4. **DNA Fragments are Treated to Make Them Single Stranded (IV)**: Following electrophoresis, the double-stranded DNA fragments are denatured to create single-stranded DNA. This is necessary for the hybridization step. 5. **Blotting (III)**: The next step is blotting, where the single-stranded DNA is transferred from the gel onto a membrane (usually nylon or nitrocellulose). This step helps to preserve the DNA fragments for further analysis. 6. **Hybridization with Radioactive Nucleic Acid (Probe) (II)**: After blotting, a labeled probe (which is a radioactive nucleic acid) is applied to the membrane. The probe will hybridize with the specific DNA sequences of interest. 7. **Autoradiography (I)**: Finally, autoradiography is performed to visualize the hybridized DNA. This step involves exposing the membrane to X-ray film, which will show the locations of the radioactive probes that have bound to the DNA. ### Correct Sequence: The correct sequence of the Southern blot procedure is: VII → VI → V → IV → III → II → I
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