Errors in Gel electrophoresis

Gel electrophoresis

Gel electrophoresis is one of the main methods applied in molecular biology for the analysis of DNA. This method includes the migration of fragments of DNA over a gel, where they are separated based on size or shape. However, even a scientifically method such as gel electrophoresis is not immune to errors and users probably face to some errors.

Introduction of Electrophoresis

Gel electrophoresis includes using a gel usually made of agarose powder (Refer to YT9059). The gel is plunged in a buffer solution leading an electric field. The DNA sample is fragmented using restriction enzymes and is injected into the gel. When the electric field is turned on, the DNA fragments in the gel move toward the positive electrode. Different sizes in DNA fragments makes migration times different due to each size fragment. The fragments are made visible using a dye or autoradiography as bands in the gel.

Contamination of the DNA Sample

The main application of electrophoresis is determined to analyze of DNA in molecular biology, but it is also used for identifying samples from a crime scene in forensics. It is important that sources of errors in this technique be minimized in order to get true results. One of the common errors is contamination of the DNA sample. If there is foreign DNA in the sample, the gel will show more extra bands in a gel .

Problems with the Gel, Current and Buffer

Another important point to avoid errors is concentration of the gel. Fragments moving is under the influence of concentration which causes moving too high or too quickly. This will be one of the reasons for errors in resolving the different bands. During the electrophoresis run, make sure that the voltage is stable. Any fluctuations in the voltage will makes unsteady migration of DNA fragments. Also buffer solution must also be noticed of the correct composition, as a buffer with the wrong pH or ionic concentration will change the shape of the DNA fragments and changing their movement times.

Proper Visualization in Gel electrophoresis

The most significant point is accurate visualization of the gel .this visualization depends on concentration of the dye or radioactive probe and makes a messy resulting image if it gets too high. Also, user doesn’t have any visualization if this concentration gets too low.

By following all correct processes during all stages, gel electrophoresis will yield accurate results. All scientific researchers should be noticed of prone to errors in gel electrophoresis, but these points can minimize errors.

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100bp DNA ladder | 50bp DNA ladder | 1kb DNA ladder

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