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Essay of gel electrophoresis
Essay of gel electrophoresis
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As seen on many crime shows and at real-life crime scenes, it is necessary to be able to identify DNA. Most of the time, this is done using a technique known as gel electrophoresis. Gel electrophoresis is a method used to separate the macromolecules that make up nucleic acids, such as DNA and RNA, along with proteins. Gel electrophoresis is significant because it has given scientists insight on what cells cause certain diseases and has led to advancements in DNA and fingerprint identification. My experiment will use gel electrophoresis to compare samples of natural and synthetic food dyes. The background for this experiment broaches the following subjects: inventors, real-world uses, necessary components, separation, and information on food dyes.
Electrophoresis was first developed in the 1930s by Tiselius. It has since expanded, and new techniques have been developed. The system of gel electrophoresis was developed in the 1950s by Oliver Smithies (Oliver Smithies: Born Inventor). Smithies created a new technique using starch and staining within gel to allow for better protein resolution. This technique was revolutionary because it was relatively inexpensive and easy to use. It provided great clarity of samples. Smithies has won many awards for his work, including the Nobel prize, and his technique is used daily by modern molecular scientists (Oliver Smithies- Biographical).
Gel electrophoresis is used in a variety of settings, particularly in molecular biology. Besides being used to separate nucleic acids, such as DNA and RNA, gel electrophoresis is also employed to divide proteins (Gel Electrophoresis). According to research, electrophoresis is applied for the following reasons, "To get a DNA fingerprint for forensic pur...
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...hat are macromolecules?" November 14, 2013. December 12, 2013. http://chemwiki.ucdavis.edu/?title=Under_Construction/Schaller/Part_I:__Structure_in_Organic,_Biological_%26_Inorganic_Chemistry/MM.__Macromolecules_and_Supramolecular_Assemblies/MM1._What_are_macromolecules%3F
Slutz, Ph. D.,Sandra. "Forensic Science: Building Your Own Tool for Identifying DNA." September 5, 2013. December 2, 2013. http://www.sciencebuddies.org/science-fair-projects/project_ideas/BioChem_p028.shtml#summary
Smithies, Oliver. " Oliver Smithies- Biographical." 2007. December 16, 2013. http://www.nobelprize.org/nobel_prizes/medicine/laureates/2007/smithies-bio.html
Unknown Author. "Monosaccharides." December 14, 2013. http://www.princeton.edu/~achaney/tmve/wiki100k/docs/Monosaccharide.html
Williams, Ruth. "Oliver Smithies: Born Inventor." Circulation Research.108.6 (2011).
Digestion of the haemolytic and non-haemolytic cells allowed for easier identification of fragments during electrophoresis analysis. Lane 12 in figure 3 show the size markers of SPP1 digested with EcoR1 while lanes 6 and 7 show samples of pK184hlyA and pBluescript digested with EcoR1 and Pst1. Lane 4 was loaded with plasmid DNA from haemolytic cells digested with EcoR1 and Pst1 while lane 5 was loaded with EcoR1 and Pst1 digested DNA from non-haemolytic cells. There was a lack of technical success in both lanes due to no bands appearing in lane 4 and only a single band appearing in lane 5. Theoretically, two bands should appear in both lanes after successful to allow for fragment identification. A possible explanation for the single, large fragment in lane 5 is that successful digestion did not take place and the plasmid was only cut at one restriction site leaving a large linear fragment of plasmid DNA. The absence of bands in lane 4 could be because there was not enough plasmid loaded into the lane. Another possibility could be that low plasmid yield as obtained when eluting the experimental samples in order to purify it. Lanes 8 and 9 belonged to another group and show technical success as two bands were present in both the haemolytic (lane 8) and non-haemolytic (lane 9) lanes. If the
The DNA retrieved from the reaction can then be. imported into an apparatus using gel electrophoresis to compare the sample of DNA to other. samples. The. In our experiment we learned the how to replicate tiny samples of DNA into usable amounts and how to analyze the specimen using gel electrophoresis.
The repeat segments are cut out of the DNA strand by a restrictive enzyme that acts like scissors and the resulting fragments are sorted out by electrophoresis (Saferstein 391). However, there are some drawbacks using the RFLP method in the forensic science community. The RFLP technique requires a large amount of DNA and must be of high quality and cannot be degraded (Jones). Forensic scientists and the law enforcement community determined a need for a DNA profiling method that could be used on smaller DNA samples. Thus, the RFLP technique has been almost entirely replaced by Polymerase chain reaction.
During this time, it could only be used in a lab with semi-intense supervision. Now, fast forward a few decades and there are D.I.Y. at home kits. The process of Electrophoresis starts with an electric current being run through a gel containing the molecules of interest. The molecules will then travel through the gel in different directions and speeds, based on their size and charge, allowing them to be separated from each other. Dyes, fluorescent tags, and radioactive labels can all enable the molecules on the gel to be seen after they have been separated. Because of these identification markers, they appear as a band across the top of the gel. Electrophoresis can be used for many different things. It is used to identify and study DNA or DNA fragments, and helps us to better understand the molecular components of both living and deceased organisms. Electrophoresis can also be used to test for genes related to specific diseases and life altering diagnoses such as Multiple Sclerosis, Down’s Syndrome, kidney disease, and some types of cancer. Electrophoresis also plays a major role in the testing of antibiotics. It can be used to determine the purity and concentration of one specific type of antibiotic or several general antibiotics at a time. Electrophoresis is also extremely useful in the creation and testing of
A DNA sample was then taken from the crime scene, Anna, and Erica to officially identify whose blood was found using gel electrophoresis. The restriction enzyme HaeIII, which cuts the DNA strand at the sequence GGCC, was placed into the samples of DNA and then ran through the gel. When the process was complete the crime scene DNA profile matched identically to Anna’s DNA profile with lines showing up at lengths 12, 23, 11, 33, and 24. After the conclusion of this experiment it was concluded that the blood found at the crime scene belonged to Anna
It was in 1984 when Alec Jeffreys, a British geneticist, discovered that specific sequences of DNA did not add to the function of a gene but were still constant throughout it. (Britannica). Jeffreys called these minisatellites and determined that each individual organism had a unique arrangement of minisatellites (Britannica). In the early uses of DNA fingerprinting, it was only used for identifying genetic diseases and disorders but people quickly realized that it could be used in many different areas of science (hubpages). Years after the discovery of DNA fingerprinting, it had been used to solve the first immigration case, the first paternity case, and even helped identify the first identical twins (le.ac.uk). The first methods of DNA fingerprinting were accurate, but you would have had needed to acquire a large amount of DNA. Over time, the advancement of science has led to major advances that formed the basis of DNA profiling techniques. These newer methods are still used today and allow scientist to use skin, blood, semen, and hair to gather DNA (le.ac.uk). In 1988 DNA fingerprinting was used for the first time in a criminal investigation. Timothy Spe...
In this experiment, the chromatography of skittles and Crayola markers will be used to determine how the dyes that are used in the markers and candy of the same color compare to each other. By finding the chromatography of these substances, we will be able to determine whether the dyes used in these materials are the same or if they differ. We will also be able to determine if the dyes used in Crayola markers are safe to digest. My hypothesis is that the dyes used in Crayola markers and skittles of the same color will differ, but the dyes used in Crayola markers are not toxic. To better support this hypothesis, the process of chromatography and the toxicity of various dyes will need to be explained.
This piece is more of a history of TMV and its uses. Modern techniques would not be appropriate. The goal is not to portray each aspect of TMV or to describe every inkling we have learned very recently. Creager uses the scientific modes of the time period in which she is describing - 1930-1965. She identifies the use of ultracentrifugation, the first isolation of TMV, and some of the first uses of electrophoresis. Each of these techniques has been greatly altered since the mid-20th century. That is not the point, however. As a History professor, it is logical for Creager to use contemporary models to evaluate other contemporary models, like
Once a crime has been committed the most important item to recover is any type of evidence left at the scene. If the suspect left any Deoxyribonucleic acid (DNA) at the crime scene, he could then be linked to the crime and eventually charged. A suspect’s DNA can be recovered if the suspect leaves a sample of his or her DNA at the crime scene. However, this method was not always used to track down a suspect. Not too long ago, detectives used to use bite marks, blood stain detection, blood grouping as the primary tool to identify a suspect. DNA can be left or collected from the hair, saliva, blood, mucus, semen, urine, fecal matter, and even the bones. DNA analysis has been the most recent technique employed by the forensic science community to identify a suspect or victim since the use of fingerprinting. Moreover, since the introduction of this new technique it has been a la...
DNA fingerprinting is a process with many steps that takes a lot of time. The first step of making a genetic fingerprint requires getting a sample of DNA, a thread of hair, a drop of blood, even skin cells. Any one of these samples of DNA contain white blood cells which are broken down by using a detergent. Thi...
Chromatography and electrophoresis are used for many wonderful processes that take place in labs. Chromatography and electrophoresis are separation techniques employed by chemists. Chemists execute multiple chemical tasks daily therefore they must find the most efficient ways to carry out these functions. Even though electrophoresis has some benefits, Chromatography is more useful than electrophoresis because it is more versatile and can be widely used in research; chromatography should be used more than electrophoresis in scientific research because it is more effective in a lab setting and can be used in more fields.
LAB REPORT 1st Experiment done in class Introduction: Agarose gel electrophoresis separates molecules by their size, shape, and charge. Biomolecules such as DNA, RNA and proteins, are some examples. Buffered samples such as glycerol and glucose are loaded into a gel. An electrical current is placed across the gel.
The use of DNA evidence was first used in a criminal investigation in Leicestershire, UK in 1986 by Professor Jeffrey, who used this technique to link two different cases together by DNA fingerprinting. He linked two semen stains from two cases, three years apart in two rape/murder cases, securing the perpetrator a man named Colin Pitchfork, and exonerated an earlier convicted seventeen-year-old teenager (Kilpatrick). Nowadays there are many cases such as this, involving DNA evidence such as restriction fragment length polymorphism (RFLP) testing and polymerase chain reaction (PCR) testing on DNA from the mitochondria of the cell. RFLP testing usually requires a sample that has 100,000 or more cells (such as a dime-sized bloodstain) and contains
He distinguished between arteries and veins as well as established embryology by studying chicks.... ... middle of paper ... ... They determined that DNA was a double helix structure composed of base pairings, with a sugar phosphate backbone. This model explained how “genes can duplicate themselves [and] would eventually lead to our current understanding of many things, from genetic disease to genetic engineering” (Salem).
This causes the DNA fragments to move through the gel depending on their sizes. With this, the DNA fragments will show a sample that will determine how large they are to one another. Gel electrophoresis uses a horizontal gel-like slab. These gels are made of polysaccharide called agarose, which is dry, powdered flakes. When the agarose is heated in a buffer, it makes the gel form solid, slightly squishy gels. (Dickey, J. L. 2012) At one end of the gel, there is square shaped space that is called wells, this is where the DNA sample will be placed. Before the DNA samples are added, the gel must be added to the well of the gel box. One end of the box is hooked to a positive electrode and the other a negative electrode. When active, the DNA samples will start to migrate from the positive to the negative showing a difference with the DNA samples with some fragments that are larger and some that are smaller. (Mader. S. S. 2016)