DNA And The Process Of Electrophoresis

The process of DNA Electrophoresis is very intricate; there are many factors that can push the experiment either way. If done right Electrophoresis can help change the world in a dramatic way. The most advanced discoveries have come from DNA Electrophoresis like: cloning, DNA Fingerprinting, and the discovery of DNA’s shape and form it takes. DNA Electrophoresis is a long but rewarding process that can determine important elements of life forms.

DNA, or deoxyribonucleic acid, is the base of life itself. It determines who and what we are; it makes our hair brown and our eyes blue. There are no two DNA strands that are the same. If you were to look for DNA in the body you would find most of it in the nucleus of the cell (there it is called nuclear DNA), and the rest of it would be in the Mitochondria. As there is DNA in the Mitochondria, it is not yours, it is your mothers. DNA is information put in a pattern by chemical bases: adenine, guanine, cytosine, and thymine. Theses bases pair up with each other A with T, and C with G. Each base is attached to a sugar molecule and a phosphate molecule, all together it is called Nucleotide. Nucleotides are two strands ordered as a spiral called a double helix. Human DNA comprises of around 3 billion bases, and in excess of 99 percent of those bases are the same in all individuals. The order, or grouping, of these bases decides the data accessible for building and keeping up a organism, similar to the way the alphabet appears when shows up in a specific order to frame words and sentences. DNA was a mystery until we could extract and find ways to do test it.

The extraction of DNA has been a pivotal point in starting research to routine analytic and remedial decision-making, it also determines the characteristics like the size and shape of DNA. DNA Extraction is the process by which DNA is separated from proteins, membranes, and other cellular material contained in the cell from which it is recovered. This extraction can be one of the most labor-intensive parts of DNA analysis (Elkins, 2013). DNA is very intricately placed and formed, as is the process of extracting it out of a cell; you have to follow multiple steps to the exact or the exaction won’t work. There are numerous ways to extract DNA, you just have to find the right procedure to the right organism. Some procedures can be done overnight in an incubator, some can be done in a few minutes, and some can be done in a few hours. When you are starting off in the procedures you have to find a way to breakdown the cell wall to get through to the nucleus. There are multiple ways to do this: mortar and pestle, blending, adding meat tenderizer, or extraction buffer. Once you break down the cell wall and get it to a slur you have to strain the solution to separate the junk from the DNA. At that point when you have strained enough solution out you need to put it in a test tube and slowing pore ice cold ethanol down the side of the tube. If you have successfully extracted DNA from your organism then there will be a snot like object making a swirling pattern in the ethanol. Now you can pipette the swirl with a micropipette and place it in a microtube and centrifuge it for three minutes on the highest RPM. After the three minutes you can take out your microtube and examine the bottom of the tube you will find a small pellet which is your DNA. The next step is to let it air dry out until there is no more liquid drops in the tube, at that point you can add fifty microliters of TE buffer, now your DNA is ready to be put in an Electrophoresis.

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DNA Electrophoresis is a lab technique used to analyze and determine different things about DNA. It’s the separation of negatively charged DNA molecules by an electric current passing through an agarose gel. Note that the larger fragments fluoresce more intensely. Although each of the fragments of a single class of molecule are present in equimolar proportions, the smaller fragments include less mass of DNA, take up less dye, and therefore fluoresce less intensely(Carr, 2012). Many labs use this method to help their cause such as: DNA fingerprinting, to tell the difference between the suspects DNA and the DNA at the crime scene, to study DNA on the molecular level, and helps prepare DNA for cloning and genetic engineering. Just like DNA being unique so is the pattern that takes place on the agarose gel whenever it’s ran through an Electrophoresis, this is called banding. The only situation that DNA could even be relatively close to being identical is from twins; even in that case the DNA strand has its own code. Though still more similar than ordinary siblings, they are no longer as identical as one may believe. These differences mean that despite their virtually identical genotype, monozygotic (MZ) twins can still be phenotypically different (Straiton, 2019). There are two pieces of equipment that make DNA Electrophoresis happen: the Power Supply and the Electrophoresis Tank. The Electrophoresis Tank has a positive and a negative charged electrode with wires leading out to the Power Supply. The Power Supply has many options to run the agarose gel through to draw the DNA out of the wells as quick or as slow as you want. The Power Supply for the Electrophoresis tank has the option to run voltage or amps, the time you would like to run it at, and the option to run, stop, and pause the Power Supply.

When you have your extracted DNA and ready for Electrophoresis you must prepare the equipment. There are many types of gels you can use but it’s best to use agarose gels, once you pull an adequate gel you set it along the bridge of the Electrophoresis Tank making sure that the wells of the gel are on the negative side. Like the gels there are many types of buffer you can use to put in the Electrophoresis but the one that minimizes overheating and problems within the Electrophoresis tank is TAE buffer, add the TAE buffer to where it barely covers the gel; if there is a overflow of buffer over the gel the DNA can be swept easier into the buffer. The wells are tiny rectangular shaped holes in the gel that you put your DNA in and from there the DNA will pass through the pores of the gel and the smaller size molecules of DNA will go farther than the bigger sized molecules. Whenever you have the DNA in the wells and everything is set up you can now put the lid on top of the tank plug in the cords from the tank lid into the Power Supply. The most effective setting to put onto the power supply is to set it on one hundred and twenty volts with twenty minutes to run it.

The human eye is unable to see the DNA after it is ran through the Electrophoresis without a dye or computer to point out the bands. The most common dye used to put the agarose gels through is Bromothymol Blue. There is multiple ways to dye it with Bromothymol blue, the most effective takes a twelve hour cycle of continues rocking of the dye through the gel. This can be done off of the Rocker which some labs have for that exact reason, the Rocker can also be used to dilute the dye out of the gels. When running the gels through the Rocker with the Bromothymol Blue you will put the gel in a weigh boat and put as much dye as it takes to make a nice thin layer over the gel so it can rock over time and slowly absorb in. When the time is over and the gel has absorbed as much dye as needed you will be able to see the bands popping out of the gel in a different shade of blue then the gel is.

As DNA is specific, it is who we are. Electrophoresis is the way to identify people and use the process to find or sometimes determine the actions of the organism. Before DNA Electrophoresis there was no fingerprinting, no DNA tests for babies, and no cloning to help grow the population of plants and animals. There has not been an invention and discovery that has changed science and life itself like Electrophoresis has. Thanks to Electrophoresis we have advanced in science tremendously.

References

1. Elkins, K. M. (2013). DNA Extraction. Retrieved November 10, 2019, from https://www.sciencedirect.com/topics/neuroscience/dna-extraction.
2. Carr, S. M. (2012). Retrieved November 10, 2019, from https://www.mun.ca/biology/scarr/Gel_Electrophoresis.html.
3. Straiton, J. (2019, August 5). Do Identical Twins Have the Same DNA? Retrieved November 10, 2019, from https://www.biotechniques.com/omics/not-so-identical-twins/.