DNA agarose gel: one than one chromosome, one band.

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In summary: If you have an insertion in your DNA, it will make a bigger difference in migration than if you don't, because the migration distance is based on the size of the chromosome, not the size of the DNA.
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nobahar
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Hello!

If I run an agarose gel of undigested DNA of an organism with more than one chromosome, and obtain one band (slightly smeared), what does this mean? The chromosomes aren't all the same length (some might be approximately half the size of others). Is it because they all become "entangled?", they run a distance shorter (i.e. suggesting they are larger) than anyone chromosome. One could even speculate that the band has migrated a distance expected for the genome size, but this might be influenced by the above thought (since the resolution at that large a size makes it difficult to tell).

Any help appreciated,
Nobahar.
 
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  • #2
Aha! I found pulse field gel electrophoresis, and on wiki it says the following: "DNA molecules larger than 15-20kb migrating through a gel will essentially move together in a size-independent manner."
So my undigested DNA moves at the same rate, apparently. EDIT: realized a part doesn't make sense so it is removed: I misinterpreted what it meant and in also in a way ignored what it meant.
This makes sense for why an organism with more than one chromosome shows one band. However, with a small insertion, the migration through the gell is retarded, this is not size-independent, then. If they move indpendent of size then an insertion would make no difference, so there would be no difference in migration between large DNA with or without an insertion. My results would make more sense if the DNA became 'entangled', that way insertions can be additive, which would account for a smalll inseertion making a large difference in migration, and at the same time explain (maybe) why there is only one band for an organism with multiple chromosomes.
Has anyone heard of multiple chromosomes becoming entangled?
Thanks in advance.
 
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  • #3
Basically chromosomes are just too big to be effectively separated with conventional gel electrophoresis, you need pulsed field gel electrophoresis, or possibly liquid chromatography with a really really long column length. Depending on how big the difference between chromosomes in basepairs is, you may need quite a long time to detect the differences in length (i.e. 100bp difference between chromosomes will take much longer to resolve than a difference of 20kbp)
 

1. What is DNA agarose gel electrophoresis?

DNA agarose gel electrophoresis is a common laboratory technique used to separate and analyze DNA molecules based on their size and charge. It involves running an electric current through a gel made of agarose, a polysaccharide derived from seaweed, which acts as a sieve to separate DNA fragments.

2. How does agarose gel electrophoresis work?

Agarose gel electrophoresis works by creating an electric field that causes negatively charged DNA molecules to move through the gel towards the positive electrode. Smaller DNA molecules move faster and therefore travel farther through the gel, while larger molecules move slower and travel a shorter distance.

3. Can agarose gel electrophoresis be used to analyze more than one chromosome?

Yes, agarose gel electrophoresis can be used to analyze more than one chromosome. Each chromosome contains multiple DNA molecules, and these can be separated and visualized as distinct bands on the gel. However, in order to analyze multiple chromosomes, the DNA must first be extracted and purified from the sample.

4. Why does agarose gel electrophoresis result in one band for one chromosome?

This is because each chromosome contains multiple DNA molecules that are tightly packed together. During electrophoresis, these molecules are separated based on size, but they still remain close to each other and form a single band on the gel. Therefore, one band on the gel represents one chromosome.

5. What can be learned from the resulting band pattern on an agarose gel?

The band pattern on an agarose gel can provide information about the size and quantity of DNA molecules present in a sample. This can be used to identify genetic variations, mutations, or changes in the DNA sequence. It can also be used for DNA fingerprinting or to confirm the presence of a specific DNA fragment.

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