DNA Fingerprinting Using Agarose Gel Electrophoresis
Introduction
Agarose gel electrophoresis is a form of gel electrophoresis that can separate a mix of DNA and proteins through agarose gel. It separates DNA by length or size through gel when an electric current is applied. Shorter fragments travel faster than long through the gel allowing for matches to be identified by similarity. The fragment length of DNA is different for each individual because sequences cut at specific sites. PCR or polymerase chain reaction is a technique that is used to replicated small amounts of DNA in a test tube. The replicated DNA is then treated with restriction enzymes, which cut the DNA at their specific sites.
Purpose
The purpose of this lab was to understand how to analyze a series of genetic data from DNA using agarose gel electrophoresis and decode the results.
Materials
Micropipette
Micropipette tips
Premade Agarose gel
Gel electrophoresis machine
DNA samples
‘Marker’ sample
UV light box
Methods
1. 1% agarose gel was made and allowed to solidify, and put in electrophoresis.
2. DNA was incubated with restriction endonuclease.
3. 20 µl of DNA was loaded into each well according to instructions.
4. Gel was run at 100 mV until dye was near the bottom of
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Skinner was the victim of a murder by pipette. The two suspects were Mr. Runyon and Mr. Bell. Based on the results of the gel electrophoresis, it can be concluded that Mr. Runyon killed Mr. Skinner, because his DNA reacted very similarly to the DNA left behind at the crime scene. Unlike Mr. Runyon, Mr. Bell’s DNA did not react like the sample left behind at the crime scene at all. Mr. Runyon’s, and the crime scenes DNA both ran about the same length up the gel, and at around the same concentration, with bands in the same locations. This shows how the DNA separated, and that both the DNA at the crime scene and Mr. Runyon’s DNA separated across the gel the same, proving Mr. Runyon is the
Figure 2 shows the results of the electrophoresis. Lanes 5 and 7 indicate the fragments obtained when the plasmids are digested with both restriction enzymes, indicating the approximate fragment size for the hlyA gene, the pK184 plasmid and the pBluescript plasmid. This is useful for identifying the recombinant DNA needed for this experiment
The two modes of analysis that will be used to identify an unknown insert piece of DNA would be plating the transformation cells onto LA plates that have either ampicillin or chloramphenicol and PCR. We will use the PCR thermocycler to denature the restriction enzymes that were specifically used to assimilate the vector DNA. It is important to use the PCR thermocycler because denaturation of the restriction enzyme will prevent the restriction enzyme from cutting the vector DNA, after the insert DNA has assimilated to the vector DNA. After the addition of specific primers that complement the base pair to its corresponding target strand, PCR will be used. Subsequently, Taq polymerase will be used to determine whether the insert DNA has been properly assimilated to the vector DNA. Within this specific situation, the target strand will be the insert DNA. After we let the PCR thermocycler run for approximately 2 ½ hours, we will then put our PCR products in the gel and run the gel to completion. After the gel has run to completion, we will then take a photograph of the gel using the UV transilluminator with the assistance of our TA. If the insert DNA was properly assimilated to the vector DNA, then our corresponding gel photo would have one band. After the cells have been transformed, we would g...
The Hillside Strangler was the name given to two killers who strangled and killed their victims, dumping their bodies in the hills of Los Angeles. These killings took place during October 1977 and February 1978 involving ten young women ranging from ages 12 to 28. With over one hundred law enforcement task force members working tirelessly to solve this case, the fear with in the community was still undeniable. It wasn’t long before the clues would come in one by one, to help detectives solve this heinous crime. Once two sets of DNA was revele on the crime scenes, police realized they had not just one but two killers. These two killers were cousins Kenneth Alessio Bianchi and Angelo Anthony Buono.
Upon completion of the experiment we were able to examine the DNA. First, the electrophorese
The given DNA ladder sample and each individual ligation samples were run on 40ml of 0.8% agarose in 1x TAE buffer for approximately sixty minutes at 110V. The appropriate volume of 6x GelRed track dye was used after it was diluted to a final concentration of 1x and incubated for thirty minutes. Finally, the gel was illuminated under UV light and analyzed.
The assumed murder weapon received improper testing, and DNA found on the knife proved unreliable. No blood was discovered on the knife
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.
... any of the DNA provided by the Vaninced victim support one report showed a piece of genetic material the penis of Steven branch but could not be linked to any victim.The penis of Steven branch that could not be linked to any victim or any defendant in the meantime our investigators were obtain DNA samples in the air cigarette butts world swabs from people who had some connection to the events is included samples from several people including Steven branches stepfather Terry Hobbs.The result of that analysis in May 2007 show that rope used to tie up Michael Moore could be associated with very hot provided a result the prosecution right after learning of it much more recent analysis by Mr. Fedora show that hair found on a tree root through Tree Stump at the crime scene could be associated with the DNA samples provided by Terry Hobbs.
However, this is a television show and, thus, an element of entertainment is present and inevitably skews the depiction accuracy of forensic science as a whole. Firstly, when Detective Benson is at the initial crime scene with Rebecca’s body, she examines the victim’s
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...
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.
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.
In the first experiment, the investigators were given four dyes, each representing a blood sample. One of which was present at the crime scene while the other three samples belonged to the three different suspects. An electrophoresis buffer (Tris-Borate-EDTA) was used in this experiment by having the running buffer placed in the electrophoresis chamber. After an agarose gel with four wells was added facing the negative end of the chamber, the investigators used micropipettes to transfer 10 µl of a sample into a well. Once the samples were loaded, negative and positive plugs were inserted into their respective inputs and the power source was turned on at 100 volts.
Afterwards, they transferred the DNA into a nylon membrane that has been incubated with radioactive probes that then attaches to the minisatellites. The nylon membrane is then exposed to X-ray film and then enabled for minisatellites to appear, a pattern is then developed which is called the “DNA fingerprint,” (DNA fingerprint). DNA in its essence, contains information that is stored as a code through chemical bases. These chemical bases are, adenine (A), guanine (G), cytosine (C), and thymine (T), which then pair up with one another, A with T, and C with G. The sequence between these bases is the information that enables for the construction of an organism (What is DNA). DNA between two individual humans, “on average, is about 99.9 percent the same,” (What is a DNA fingerprint). However, there exists within DNA, minisatellites, which are sequences of highly variable DNA that have a pattern unique to each individual. Jeffreys and his team determined through the comparison of the two semen samples that was there was enough similarity to conclude that there was a match and that both crimes were committed by the same
Modern technology allows DNA to be extracted from even the tiniest biological sample, which may be a few droplets of blood or a hair follicle. Moreover, once the samples are compiled, they can be analysed to create DNA profiles that can aid in identifying a specific person. If all tested profiles match, the samples come from the same person. Conversely, if they do not match, that means the samples come from different