Troubleshooting DNA Gel Analysis: Standard Curve and Smallest Band Size Queries

In summary, the individual ran a gel for restriction digestion using lambda HindIII as the marker. When drawing the graph on semi-log paper, the standard curve did not appear as a straight line, but rather a linear curve. They are unsure if this is acceptable. They also have a smaller digit on their semi-log graph paper for DNA size, but have a 0.5 band that they cannot accurately plot. They are confused as the distance from the well measured should be proportional to the log10 value of the DNA size, but their curve does not seem to reflect this. They are seeking assistance to resolve these issues.
  • #1
biochemist
11
0
Hi everyone, I ran a gel for restriction digestion and used lamda HindIII as the marker. When I'm drawing the graph on a semi-log paper, size on y-axis and distance from the well on the x axis, the standard curve isn't a straight line, but a linear curve. Is it fine?

Another question: the smallest digit given on a semi-log graph paper for DNA size is 1, but I've got a 0.5 band and couldn't point it out on the graph. What shall I do? Shall I just ignore this?

Very much appreciated if you could help me solve my problems
 
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  • #2
I'm really confused, cos I've learned that the distance from the well measured should be proportional to the log10 value of the DNA size. And by theory I should have got a straight line or nearly straight, but the curve I've got doesn't seem to be straight at all!
 
  • #3
Anyone can answer my question?
 

Related to Troubleshooting DNA Gel Analysis: Standard Curve and Smallest Band Size Queries

1. What is DNA gel analysis and why is it important?

DNA gel analysis is a laboratory technique used to separate and analyze DNA fragments based on their size. It is an important tool in molecular biology research as it allows scientists to visualize and compare different DNA samples, such as DNA from different individuals or species, and to identify genetic variations.

2. How is DNA gel analysis performed?

DNA gel analysis involves several steps. First, DNA samples are loaded onto a gel made of a porous material, such as agarose. An electric current is then applied, causing the DNA fragments to move through the gel towards the positive electrode based on their size. After the gel run is completed, the gel is stained with a dye, such as ethidium bromide, which allows the DNA bands to be visualized under UV light.

3. What is the purpose of using a DNA ladder in gel analysis?

A DNA ladder, also known as a molecular weight marker, is a set of DNA fragments of known sizes that is loaded onto the gel alongside the samples. It is used as a reference to determine the size of the DNA fragments in the samples. This allows for more accurate analysis and comparison of DNA samples.

4. What factors can affect the results of DNA gel analysis?

There are several factors that can impact the results of DNA gel analysis. These include the concentration and purity of the DNA samples, the voltage and duration of the electric current, and the type of gel used. It is important for scientists to carefully control these variables to ensure accurate and reproducible results.

5. What are the limitations of DNA gel analysis?

While DNA gel analysis is a valuable technique, it does have some limitations. It is limited in its ability to separate DNA fragments that are similar in size, and it may not be able to detect small differences in DNA sequences. Additionally, DNA gel analysis cannot provide information about the function or expression of the DNA fragments, only their size and quantity.

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