Gel Electrophoresis of DNA fragments produced by apoptosis

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In summary, gel electrophoresis is a laboratory technique used to separate and analyze molecules based on their size and charge. In the study of apoptosis, it is specifically used to analyze and visualize smaller DNA fragments produced during cell death. These fragments have distinct characteristics, such as a ladder-like pattern on a gel, and can provide valuable information about the extent of cell death in a sample. Agarose gel is commonly used for this type of gel electrophoresis, but there are limitations in using this technique, such as the inability to quantify DNA fragments and the time-consuming nature of preparing samples for analysis.
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Homework Statement
One mechanism involved in apoptosis is the digestion of DNA by DNAases. DNA gets digested into fragments of lengths equal to multiples of the distance between nucleosomes.

The figure attached shows the results of separation by gel electrophoresis of the DNA released by the action of DNAase.

Suggest how the pattern in the left-hand column would change if very high concentrations of DNAase were applied to the cells.
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gel electrophoresis
Electrophoresis.png


The answer is that there would only be a 250 bp fragment, and it would be brighter.

I think that there would just more fragments of all sizes, as more DNA is degraded. I don't understand why there wouldn't be fragments of larger sizes. Thanks!
 
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Not knowing anything about DNA digestion, but just applying some logic: if there is a fragment longer than the single distance between nucleosomes, what will DNAase do to it?
 
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...or if you chew your peanuts long and vigorously enough you get a mouthful of peanut butter. Not more chunks of peanuts.
 
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1. What is gel electrophoresis?

Gel electrophoresis is a laboratory technique used to separate and analyze DNA fragments based on their size and charge. It involves running an electric current through a gel matrix, causing the DNA fragments to migrate towards the positive electrode. The smaller and more negatively charged fragments will move faster and travel farther through the gel, while larger fragments will move slower and stay closer to the starting point.

2. How is gel electrophoresis used to study apoptosis?

Gel electrophoresis can be used to study apoptosis, or programmed cell death, by separating and analyzing DNA fragments produced during this process. Apoptosis results in the fragmentation of DNA into smaller pieces, which can be visualized and measured using gel electrophoresis. This technique allows researchers to study the timing and extent of apoptosis in different cell types and under various conditions.

3. What is the purpose of staining the gel during gel electrophoresis?

Staining the gel during gel electrophoresis allows for the visualization of DNA fragments. Ethidium bromide is a commonly used stain that binds to DNA and fluoresces under UV light, making it easier to see the separated fragments. This allows researchers to analyze the size and quantity of DNA fragments produced by apoptosis.

4. How can gel electrophoresis results be interpreted?

The results of gel electrophoresis can be interpreted by comparing the size and position of the DNA fragments to known standards or markers. This allows for the determination of the size of the fragments and the estimation of the extent of apoptosis. Additionally, the intensity of the bands on the gel can provide information about the quantity of DNA fragments present.

5. What are the limitations of gel electrophoresis in studying apoptosis?

While gel electrophoresis is a useful technique for studying apoptosis, it has some limitations. It can only provide information about the size and quantity of DNA fragments, but not the specific genes or sequences involved in the process. Additionally, it is a time-consuming and labor-intensive technique, and the results can be affected by experimental conditions such as the concentration of DNA and the voltage used during electrophoresis.

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