How to make and run an agarose gel (DNA Electrophoresis) - Article Example

Agarose Gel Electrophoresis of DNA For isolation of genomic DNA, agarose gel electrophoresis is done which is based on the principle that on disruption and lysis of cell components, followed by removal of proteins and sugar, nucleic acids can be recovered which can be observed through a UV transilluminator on the agarose gel due to prior staining by ethidium bromide.Materials: Agarose powder, Electrophoresis buffer (TAE), gel casting tray, Ethidium bromide, Comb, Sample, Loading buffer, UV transilluminator.

Method: Agarose solution (generally .8%) is prepared for isolation of DNA. However, the concentration depends upon the sample i.e. the size of DNA. Appropriate amount of agarose powder is weighed and mixed with electrophoresis buffer. The two are mixed properly and heated till the solution is clear. The solution is then left to cool till it reaches 60C. Then 1µLof ethidium bromide is added to the solution and mixed properly. This helps in observing the sample against the UV transilluminator.

The solution is poured in a gel casting tray, the two open sides of which have been covered with a cellotape. Place a comb lightly at one end and the tray is left as such to allow the gel to set. When the gel is set the sample can be loaded in the wells. The sample is mixed with the loading buffer, approximately .2 volumes of it and loaded in the well. The first well can be loaded with a marker, which can be used to determine the size of the DNA sample. The gel is run at 50V for 45 min.

The gel is observed under UV transilluminator to check the sample.Result: http://www.methodbook.net/dna/agarogel.html