Agarose gel electrophoresis of DNA is used to seperate mixtures of protein and nucleic acid fragments. Molecules tend to differ in size, charge, and mobility. The objective of this procedure is to predict the distance of DNA migrating towards the positive electrode in the electrophoresis chamber. Many scientists use agarose gel electrophoresis to examine the DNA of criminal suspects.
The first step in this procedure is to prepare a buffer containing 1% of the liquid agarose gel. Once the gel solidifies within 15 to 20 minutes, a mold should have formed. Next, a comb is inserted into the chamber to seperate the gel creating tiny holes called “wells”. The comb is removed carefully so it will not disrupt the wells inside the chamber. The DNA prepared can be loaded into the buffer using a micropipette. The DNA is added to …show more content…
Bubbles are an indication that the sample is on and running. DNA is observed from a distance to estimate how far the nucleic acid fragments have traveled based on the flurescent dye. These fragments are stained bromophenal blue or xylene cyanal. Once migration has occurred, the DNA fragments are stained with ethidium bromide. Ethidium bromide is very toxic and can cause mutations. You should always wear gloves to protect yourself, when handling these gels or solutions. This dye causes the DNA’s mass and rigidity to be altered and therefore its mobility to change. The mobility of DNA is determined by the ionic strength of the buffer supplied. There are several buffers that are used for this procedure to allow for DNA to be monitored in the gel. One of the electrophoresis buffers is EDTA or ethylenediaminetetraacetic
The first step in this procedure is to prepare a buffer containing 1% of the liquid agarose gel. Once the gel solidifies within 15 to 20 minutes, a mold should have formed. Next, a comb is inserted into the chamber to seperate the gel creating tiny holes called “wells”. The comb is removed carefully so it will not disrupt the wells inside the chamber. The DNA prepared can be loaded into the buffer using a micropipette. The DNA is added to …show more content…
Bubbles are an indication that the sample is on and running. DNA is observed from a distance to estimate how far the nucleic acid fragments have traveled based on the flurescent dye. These fragments are stained bromophenal blue or xylene cyanal. Once migration has occurred, the DNA fragments are stained with ethidium bromide. Ethidium bromide is very toxic and can cause mutations. You should always wear gloves to protect yourself, when handling these gels or solutions. This dye causes the DNA’s mass and rigidity to be altered and therefore its mobility to change. The mobility of DNA is determined by the ionic strength of the buffer supplied. There are several buffers that are used for this procedure to allow for DNA to be monitored in the gel. One of the electrophoresis buffers is EDTA or ethylenediaminetetraacetic