Gel Electrophoresis Research Paper
Restriction Enzymes and Electrophoresis
Bhumik Patel
Phillips
1/16/11
Restriction enzymes are tools in DNA research that can cut DNA into exactly needed pieces. Certain cuts can be rough, while others can be clean. Certain cuts can have an organized pattern to have a staggered cut. Other cuts will leave complementary bases with them. Electrophoresis allows the manipulation of DNA to separate and organize those parts. Electrophoresis is the substrate electric movement of the separation of DNA. Gel Electrophoresis can separate both DNA and RNA. This can also allow the knowledge of the unknown bases of a strand of DNA. This can allow the knowledge of familiar patterns that can create a unique signature. Separation of these molecules …show more content…
depends on the charge of the molecule and the mass. The charges are shown toward the opposite charge and it will attract to. The gel that organizes the DNA fragments is agarose and polyacrylamide. Polyacrylamide is a material that is similar that is found in soft contact lenses. DNA fingerprinting is a way of showing differences in DNA from one being to another. 99% of the human DNA is similar to all other humans. The remaining 1% is enough to tell us the differences from one to another. The one-percent has a very large possibility of options for your DNA base sequence. We were to determine that electrophoresis can show differences in DNA. We had used in this lab 0.8% agarose gel, gel tray, TBE Running Buffer 1X (350ml), DNA Strain, Staining Tray, Micropipets, Metric Ruler, semi-log graph paper, goggles, aprons, and gloves. We shared all the DNA samples. Molding the gel: 1) Add rubber stoppers to the ends of the gel plate 2) Add well placements 3) Add 25ml of agarose gel to gel plate 4) Add gel stain 5) Let the gel rest for about 20 minutes or until opaque.
Loading and running the gel: 1) Remove rubber stoppers and well placement from the gel plate. (remove gel placement straight up) 2) Add 10µl of each DNA sample to a well in the gel. (Avoid piercing the bottom of the wells with the tip of the micropipette tip.) 3) Add 350 ml of 1X TBE running buffer to the electrophoresis unit. 4) Place loaded gel into electrophoresis unit. 5) Attach cover, make sure the patch cords are connected and dry, and plug in the red patch cord onto the red electric slot, do the same for the black patch cord. 6) Plug in electric supply. 7) Turn on the electric supply; look for a red light on the power supply and bubbles along the platinum electrodes. 8) After given time, turn off the power supply and remove the gel plate. 9) Put gel (with or without gel plate) under a light box or black light box and look at the results.
Analysis:
1) Measure each DNA bands, in millimeters. Measure from each bottom of each well to the center of each DNA strand. 2) Record the measurements. 3) Graph the results on semi-log graph paper to compare results.
DNA Marker Standard (Lambda DNA/Hind III Digest) (in pencil) Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 23,130 | 2 | 7 | 9,416 | 3 | 8 | 6,557 | 4 | 10 | 4,361 | 5 | 14 | 2,322 | 4) Using this graph we are able to determine the lengths of the other segments.
Crime Scene DNA Sample Fragment | Distance Migrated (mm) | Length (bp) (all are approx.) | 1 | 6 | 9,400 | 2 | 10.5 | 5,600 | 3 | 15 | 3,275 | 4 | 22 | 1,475 | 5 | 27 | 820 |
Suspect 1 DNA Sample Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 9,400 | 2 | 10 | 5,825 | 3 | 12 | 4,700 |
Suspect 2 DNA Sample Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 9,600 | 2 | 10.5 | 5,600 | 3 | 16 | 3,275 | 4 | 22 | 1,475 | 5 | 27 | 820 |
1.
Compare the banding patterns formed in each lane of the gel. Assume it is the same as the banding pattern on an autoradiograph. Do you think the three DNA samples tested are the same? Explain. How can you further verify weather or not any of the DNA samples tested are the same?
A: No, because the samples would lead to, if they are all the same, that all of the suspects have committed the crime or that they are identical twins. You could have done a lie detector test as a further verifier. 2. Which of the two suspects if the real burglar? Explain you answer.
A: The second suspect is more likely to be the burglar because the DNA matches more on the second suspect than the first one. 3. Assume you have to identify two unknown DNA samples. Explain the function of each step in the DNA fingerprinting technique. a) Restriction Enzyme Digestion cleaves and cuts DNA in certain spots in to fragments. b) Gel Electrophoresis can be used to separate and organize the DNA fragments. c) Denaturation into Single-Stranded DNA this will divide those fragments in to more distinct categories and fragment
size. d) Southern Blotting shows DNA in size factions from nucleotide. e) The use of Radioactive DNA Probe allows us to read and see individually each fragment though a patterned sequence. f) When making an autoradiograph the lengths will show a pattern in which you would be able to show on an autoradiograph with vertical columns. 4. The application of DNA fingerprinting technology in forensic science as well as in medicine has raised many legal and ethnic concerns. Discuss the following issues: Should data banks be established for DNA information? What are some benefits of DNA data banks? How will these be controlled? Should the DNA taken from a suspect for identification be used to determine other genetic characteristics?
A: No, data banks should not be established for storing DNA information. DNA data banks would not need the need for the long testing is a great benefit. They would need to be controlled by a high security and would need to be controlled the government. Yes, DNA should be taken from suspects for the fact that their DNA should change.
The purpose of the lab was to determine that who had committed the crime as well as knowledge of how restrictive enzymes and electrophoresis can show patterns in DNA. We are to conclude that suspect 2 was the burglar because the 1st, 2nd, 3rd, 4th, and 5th fragments for the crime scene and the suspect matched exactly according to the lab. There may have been errors in how much buffer and power levels in the electrophoresis unit. There could have also been and error in how long we may have left the gel inside the unit. The long we may have had to have left it in there we may have had better results.
Bhumik Patel
Phillips
1/16/11
Restriction enzymes are tools in DNA research that can cut DNA into exactly needed pieces. Certain cuts can be rough, while others can be clean. Certain cuts can have an organized pattern to have a staggered cut. Other cuts will leave complementary bases with them. Electrophoresis allows the manipulation of DNA to separate and organize those parts. Electrophoresis is the substrate electric movement of the separation of DNA. Gel Electrophoresis can separate both DNA and RNA. This can also allow the knowledge of the unknown bases of a strand of DNA. This can allow the knowledge of familiar patterns that can create a unique signature. Separation of these molecules …show more content…
depends on the charge of the molecule and the mass. The charges are shown toward the opposite charge and it will attract to. The gel that organizes the DNA fragments is agarose and polyacrylamide. Polyacrylamide is a material that is similar that is found in soft contact lenses. DNA fingerprinting is a way of showing differences in DNA from one being to another. 99% of the human DNA is similar to all other humans. The remaining 1% is enough to tell us the differences from one to another. The one-percent has a very large possibility of options for your DNA base sequence. We were to determine that electrophoresis can show differences in DNA. We had used in this lab 0.8% agarose gel, gel tray, TBE Running Buffer 1X (350ml), DNA Strain, Staining Tray, Micropipets, Metric Ruler, semi-log graph paper, goggles, aprons, and gloves. We shared all the DNA samples. Molding the gel: 1) Add rubber stoppers to the ends of the gel plate 2) Add well placements 3) Add 25ml of agarose gel to gel plate 4) Add gel stain 5) Let the gel rest for about 20 minutes or until opaque.
Loading and running the gel: 1) Remove rubber stoppers and well placement from the gel plate. (remove gel placement straight up) 2) Add 10µl of each DNA sample to a well in the gel. (Avoid piercing the bottom of the wells with the tip of the micropipette tip.) 3) Add 350 ml of 1X TBE running buffer to the electrophoresis unit. 4) Place loaded gel into electrophoresis unit. 5) Attach cover, make sure the patch cords are connected and dry, and plug in the red patch cord onto the red electric slot, do the same for the black patch cord. 6) Plug in electric supply. 7) Turn on the electric supply; look for a red light on the power supply and bubbles along the platinum electrodes. 8) After given time, turn off the power supply and remove the gel plate. 9) Put gel (with or without gel plate) under a light box or black light box and look at the results.
Analysis:
1) Measure each DNA bands, in millimeters. Measure from each bottom of each well to the center of each DNA strand. 2) Record the measurements. 3) Graph the results on semi-log graph paper to compare results.
DNA Marker Standard (Lambda DNA/Hind III Digest) (in pencil) Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 23,130 | 2 | 7 | 9,416 | 3 | 8 | 6,557 | 4 | 10 | 4,361 | 5 | 14 | 2,322 | 4) Using this graph we are able to determine the lengths of the other segments.
Crime Scene DNA Sample Fragment | Distance Migrated (mm) | Length (bp) (all are approx.) | 1 | 6 | 9,400 | 2 | 10.5 | 5,600 | 3 | 15 | 3,275 | 4 | 22 | 1,475 | 5 | 27 | 820 |
Suspect 1 DNA Sample Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 9,400 | 2 | 10 | 5,825 | 3 | 12 | 4,700 |
Suspect 2 DNA Sample Fragment | Distance Migrated (mm) | Length (bp) | 1 | 6 | 9,600 | 2 | 10.5 | 5,600 | 3 | 16 | 3,275 | 4 | 22 | 1,475 | 5 | 27 | 820 |
1.
Compare the banding patterns formed in each lane of the gel. Assume it is the same as the banding pattern on an autoradiograph. Do you think the three DNA samples tested are the same? Explain. How can you further verify weather or not any of the DNA samples tested are the same?
A: No, because the samples would lead to, if they are all the same, that all of the suspects have committed the crime or that they are identical twins. You could have done a lie detector test as a further verifier. 2. Which of the two suspects if the real burglar? Explain you answer.
A: The second suspect is more likely to be the burglar because the DNA matches more on the second suspect than the first one. 3. Assume you have to identify two unknown DNA samples. Explain the function of each step in the DNA fingerprinting technique. a) Restriction Enzyme Digestion cleaves and cuts DNA in certain spots in to fragments. b) Gel Electrophoresis can be used to separate and organize the DNA fragments. c) Denaturation into Single-Stranded DNA this will divide those fragments in to more distinct categories and fragment
size. d) Southern Blotting shows DNA in size factions from nucleotide. e) The use of Radioactive DNA Probe allows us to read and see individually each fragment though a patterned sequence. f) When making an autoradiograph the lengths will show a pattern in which you would be able to show on an autoradiograph with vertical columns. 4. The application of DNA fingerprinting technology in forensic science as well as in medicine has raised many legal and ethnic concerns. Discuss the following issues: Should data banks be established for DNA information? What are some benefits of DNA data banks? How will these be controlled? Should the DNA taken from a suspect for identification be used to determine other genetic characteristics?
A: No, data banks should not be established for storing DNA information. DNA data banks would not need the need for the long testing is a great benefit. They would need to be controlled by a high security and would need to be controlled the government. Yes, DNA should be taken from suspects for the fact that their DNA should change.
The purpose of the lab was to determine that who had committed the crime as well as knowledge of how restrictive enzymes and electrophoresis can show patterns in DNA. We are to conclude that suspect 2 was the burglar because the 1st, 2nd, 3rd, 4th, and 5th fragments for the crime scene and the suspect matched exactly according to the lab. There may have been errors in how much buffer and power levels in the electrophoresis unit. There could have also been and error in how long we may have left the gel inside the unit. The long we may have had to have left it in there we may have had better results.