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Updated: Jul 5, 2026
07:10
Agarose Gel Electrophoresis for the Separation of DNA Fragments
Published on: April 20, 2012
Agarose gel electrophoresis
1Iowa State University, Ames, Iowa, USA.
Current Protocols in Neuroscience
|April 23, 2008
Summary
This study details a protocol for isolating and purifying DNA fragments ranging from 0.5 to 25 kilobases (kb). It also covers using midigels and minigels for DNA fragment analysis.
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- Accurate separation and purification of DNA fragments are crucial for various molecular biology applications.
- Existing methods may have limitations in terms of fragment size range or efficiency.
Purpose of the Study:
- To present a robust protocol for the separation and purification of DNA fragments.
- To describe the application of midigels and minigels within this protocol.
Main Methods:
- The protocol involves specific steps for DNA fragment isolation.
- Support protocols detail the use of midigel and minigel electrophoresis for fragment separation.
Main Results:
- The protocol enables the purification of DNA fragments within the 0.5 to 25 kb size range.
- The use of midigels and minigels facilitates efficient separation of these fragments.
Conclusions:
- This protocol provides a reliable method for obtaining purified DNA fragments of various sizes.
- The described techniques are valuable for researchers in molecular biology and genetics.
Related Concept Videos
02:35DNA Agarose Gel Electrophoresis
Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
01:22Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
01:27SDS-PAGE
Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
01:20Electrophoresis: Overview
Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
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02:57Southern Blot
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...