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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 purpose of this lab is to learn how to properly conduct two different macromolecules test, the nucleic test and protein test in order to identify whether four different types of food, contain proteins and nucleic acid. The way an individual can determine if a specific macromolecule is present is by conducting qualitative tests, which allows an individual to determine whether a certain macromolecule is present by observing the color change. Additionally, for statistical analysis semi-quantitative tests will be conducted as well to determine the relative amount of a macromolecule that is present in the food based on the color change. (Dooley 20). Moreover, before conducting this experiment an individual must determine the positive and negative
Upon completion of the experiment we were able to examine the DNA. First, the electrophorese
Many things have impacted both the Science and Medical fields of study. Electrophoresis and DNA Sequencing are two of these things. Together they have simultaneously impacted both of these fields. On one hand, there is Electrophoresis. Electrophoresis is a specific method of separating molecules by their size through the application of an electric field. It causes molecules to migrate at a rate and distance dependent on their size. On the other hand, there is DNA Sequencing. DNA Sequencing is a technique used to determine the exact sequence of bases
Polymerase Chain reaction (PCR) is used to isolate a predetermined strand of DNA on the double helix. Once the desired DNA is isolated it is able to be copied as much as needed (2). In this experiment PCR was used to isoloate Vangl2 from Zebra fish embryos. In a PCR experiment, a primer is used to find and isolate the desired nucleotide sequence of DNA (2). In this experiment two primers were used as follows:
It was loaded into the gel instead of in the well leading to no results. There should have been a series of bands similar to well three. In lanes three, 5, and 6 we see the bands labeled A,B and C. Due to the short distance traveled down can discern that these bands are in fact DNA as it is both bulky and negatively charged. In lane seven one can see the band in the beginning, which is identified, as tRNA. E and F represent the left of “junk” from the inside of the cell that made it’s way into the sample such as mRNA and proteins. This can be told by seeing that the small materials traveled very far down the gel and were not removed by DNase. What truly tells one whether or not he or she extracted DNA are the blank spots on the gel. In lane four and eight there are missing bands. This is due to the fact in these samples enzymes where added to break down the nucleic acids, DNase in the case of lane four and RNase in eight, thus causing a gap where they should appear. The data that was collected seems to indicate that the sample that was extracted was done properly and yielded DNA. It should be noted that the lanes three and four were switched when adding the material into
After extracting, filtering, and condensing the blue and pink dyes off of the candies, it can be determined that the Blue candy is FD & C Dye Blue 1 and the Pink candy is a mix of Yellow 5 and Red 40. During the solvent evaporation for the blue dye, it was observed that when concentrated, the transparency of the dye was similar to that of Blue 1. During the paper chromatography of Project 2 Session 1, the blue dye showed on the chromatogram to have the highest spot height. Similar to the paper chromatography today which the blue dye was observed having the higher spot height as well. Figure 3 of the Project 2 In-Lab Report displays Blue 1 of having an Rf value of .729. The blue dye off of the candy, as seen in Figure 2, has an Rf value of .657.
The material and equipment used was a sample of candy such as M&M’s, skittles, and Reese’s pieces. Set food colors for comparison. Filter paper or coffee filters. 0.1% salt solution {1/8 tsp salt in 3 cups of water}. Clear plastic 9 oz cups. Blow dryer. Also you will need some toothpicks and small {1 oz} plastic cups. This are the materials and equipment we used for this experiment .the objective of the experiment is to use the technique of paper chromatography to show that it can be used to separate from each other in a mixture. To understand the principles of paper chromatography.
The DNA Isolation experiment was to examine the extraction of DNA from fruits. Three fruits were used to extract DNA using different solutions. DNA was found in only one fruit among the three because of various reasons, including inaccuracy during the experiment. This experiment showed the process of how to extract DNA from organisms that could be used for PCR. The PCR stimulation was to investigate the criminal behind the assault case using gel electrophoresis. In order to find the criminal between two identical brothers, blood samples from both parties were collected for DNA fingerprinting. The purified DNA of the two suspects were placed in agarose gels and electrophoresis was performed to determine who was guilty. The evidence that matched up to the suspect was the criminal. This experiment showed how DNA fragments are separated from each other depending on size and charge.
Ballantyne, Jack, George Sensabaugh, and Jan Witkowski. DNA Technology and Forensic Science. New York: Cold Spring Harbor Laboratory Press, 1989.
Woolfe, M., & Primrose, S. (2004). Food forensics: using DNA technology to combat misdescription and fraud. Trends in Biotechnology, 22, (5), 222-226.
To begin with this project, a gel electrophoresis chamber must be built. In this chamber, a plastic box will be the chamber, a stainless steel wire will replicate electrodes, batteries will be the power outlet, and the wells will be replicated by using a styrofoam comb. Hold the plastic box horizontally.
This type of method is used in the separation and viewing of macromolecules (Wiley, n.d.). In an investigation process, a forensic analyst can perform this technique to find out if the DNA found at a crime scene is similar to or the same as the suspect's DNA (Wiley, n.d.). Another technique that forensic scientists use effectively is the liquid-liquid extraction method. With this method, drugs can be extracted from biological samples, such as blood, urine, or sweat. A variety of different drugs and metabolites can be obtained through this method. This is one way how forensic analysts are able to separate drugs. In class, we performed a lab for Week 2 which involved the extraction of caffeine from tea bags, which is a metabolite, and this was carried out through the process of the liquid-liquid extraction. This is an example of how forensic scientists apply these tests to real-life analysis of evidence in an investigation. The first part of the lab consisted of the disintegration of caffeine into
In fact, 1 Nano gram or less can produce unexceptional information to find a suspect or a victim of a crime. By means of this technology an investigator can extend their criminal investigation beyond the realms of traditional semen or dried up blood. Crime scenes may consist of less valuable DNA, it subsequently depends on the environmental conditions where the DNA was found. For certain, a sequence of factors ramifies the capability to obtain a pleasing DNA profile. By any means, extended exposure to any type of biological evidence can degrade the DNA, and therefore renders it useless for further analysis.
Agarose gel electrophoresis separates molecules by to 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 current moves the molecules towards the cathode or anode. The speed of the moving molecules depends on the size, shape, and charge. The properties of the gel will definitely affect the movement. Small molecules are expected to move easily and faster thru the pores.