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Gel Electrophoresis Lab

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Agarose gel electrophoresis is an easy and common technique of separating and analyzing DNA. The main objective of this lab is to find the sire of the offspring using gel electrophoresis. Gel electrophoresis is used in laboratories to isolate charged molecules like DNA, RNA, and particular proteins according to their specific size. The charged molecules travel through the gel when an electric current is spread across it. The electric current is applied across the gel so that the ends of the gel have a positive charge and the other end has a negative charge. Migration, in gel electrophoresis, is the movement of charged molecules. The charged molecules migrate towards its opposite charge. A molecule like DNA with a negative charge will be drawn towards the positive end. Because DNA and RNA are negatively charged molecules, they …show more content…

The gel consists of a penetrable matrix through which molecules can travel when triggered by an electric current. Smaller molecules migrate through the gel more quickly and consequently travel further. Larger fragments that migrate slowly travel a shorter distance. The result is that the specific molecules are separated by size in electrophoresis. A short tandem repeat (STR) in DNA is a quantity of polymorphisms that occurs when a pattern of two or more nucleotides is repeated and the recurrent sequences are directly adjacent to each other. Short tandem repeats are also called microsatellites. STR’s are considered to be small genetic markers found in genomic DNA that consists of two to six base pair sequence that is repeated one after the other five to 100 times. STR’s are used to determine paternity by matching STR with offspring. If the STR is not from the mother, then is has to be from the father. A lab instructor provides a micropipette, which is used to extract given DNA

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The same region is also amplified on both chromosomes, however they are different sizes, which are then put into gel

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Once the program was run, the machine began its thirty-five PCR cycles. Each step in these three step cycles is defined by a specific time span and temperature. However, before the first cycle begins, initial DNA denaturation occurred for five minutes at a temperature of 95 oC in order to ensure that all the DNA had become single-stranded. After this initial denaturation, the thirty-five PCR cycles could begin, first with thirty seconds of DNA denaturation at 95 oC, then followed by a drop of temperature to 58 oC for thirty seconds for the primers to anneal to its target sequence on the template DNA, and finally thirty seconds at 72 oC for primer extension in which DNA synthesis is carried out by Taq Polymerase. After the thirty-five cycles were complete, there was an additional five minutes at 72 oC for the final extension of the DNA to ensure that all synthesis had been completed.

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In order to do this a PCR was set up. A PCR tube was obtained from the instructor. The PCR bead was then dissolved in 22 microliters primer cocktail. After the PCR bead was dissolved, three microliters of the DNA extracted from the first part of the experiment were added to the PCR mixture. This PCR mixture and the 1.5 ml extracted DNA samples were then submitted to the

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It is an analytical method where in a protein sample is electrophoresis on an SDS- PAGE and electro transferred on the nitrocellulose membrane. The transferred protein is detected using specific primary enzymes labeled antibody. Antibodies bind to specific sequences of amino acids, known as the epitope. Because amino acid sequences are different from protein to protein, antibodies can recognize specific proteins among a group of many. Therefore, a single protein can be identified in a cell lysate that contains thousands of different proteins and its abundance quantified through western blot

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Figure 2a Figure 2b Figure 2a represents the gel fully run through gel electrophoresis for thirty minutes and figure 2b represents the expected results from the gel electrophoresis run. As seen by figure 2b, there are bands present in all lanes except for land three which is the wild type digested DNA sample. It is very faint, but in lane three of figure 2a, there appears to be a slight band apposed to the

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The key enzyme in the process is DNA polymerase that forms the PCR product by linking individual nucleotide bases (adenine, guanine, cytosine and thymine). Short DNA fragments with sequence complimentary to target DNA are called primers; these specify the DNA sequence that has to be amplified. The PCR reaction mixture is made in a 96 well plate or test tube and placed in a thermal cycler. The thermal cycler is a machine used to amplify DNA by altering the temperature in repeated cycles using three precise pre programmed steps: Denaturation – By increasing the temperature in the thermal cycler the DNA is denatured creating separate single stranded DNA

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DNA analysis could also be used in paternity cases to find out who the father/mother is. The child will have half the bands from the mother the other half from the father. Since the evidence was individual, the DNA matched only one person, Hector Hawk. This evidence is useful in the case because DNA from blood does not randomly get to the crime scene without the person being there. The blood was left behind after the victim was killed and cannot follow Locard's exchange principle because the blood is a liquid and cannot simply be traveled or exchanged by

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They apply different temperatures which results in melting of the DNA. After this process the double helix will separate and enzymatic replication will occur. Complementary primers are added resulting in multiplying of the sample. Therefore this is how Forensics investigators successfully tested the blood stain on the suspect jacket and they were able to say if Dobson was guilty for Stephen Lawrence death. DNA was first used in UK for an emigration case.

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Abstract This experiment showed that temperature, concentration and pH all affect the rate of enzyme reaction differently. Enzymes are very important in organisms and therefore understanding how and why they work the way they do in specific conditions is crucial. The results showed that an increase in temperature would also increase the reaction rate, until a temperature that was too high, where the enzymes began to denature and therefore the rate of reaction was slowed down. As concentration was increased, the reaction rate continued to increase.

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Although, in our lab report, suspect ones DNA matched the crime scene when cut with enzyme one, this can be explained by how closely related the two suspects are. Therefore two enzymes were used to cut the DNA; the suspect has to match both. Moreover, the limitation to DNA fingerprinting is, if a person were to have an identical twin. This is because identical twins have the same DNA because they come from the same egg. If a suspect’s DNA matched that of the one being tested, and they had an identical twin, a farther investigation would need to be done.

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The membrane and nucleus had to be broken down since the DNA is found inside, which is found inside the membrane. A possible source of error could have been if a chunk of a strawberry got into the test tube with the filtrated strawberry juice and DNA extraction buffer. DNA is soluble, but not in ethanol. This is why when the ethanol was added, the DNA was then visible. This could have affected the data because there might have been organelles and other parts of the strawberry besides the DNA that wasn’t supposed to be there in the filtrated strawberry juices and extraction buffer.

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Retrieving this amount of data is both exhausting and time consuming. A short cut has been found that scientist use to analyze smaller segments of DNA. Short tandem repeats (STR) are segments of DNA sequences that are repeated. (BIO) The span of each STR differs from one person to the next.

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Lesly Rodriguez Biology January 23, 2018 DNA is a long skinny molecule made up of nucleotides; the nucleotides consists of adenine, thymine, cytosine, and guanine. DNA also carries genetic information in cells and some viruses which forms two long lines of nucleotides twisted like a zipper known as the double helix. DNA can be extracted and kept in a database from having samples such as hair, saliva, fingertips, etc; the purpose of keeping it in a database is in order to secure vital information of an individual’s privacy and use it for an important event. The city of Chicago should make it mandatory for individuals and newborn babies to submit their DNA into a database. I foresee this due to solving crime efficiently, knowing circumstances that are from relatives, and the scientific method cures for the circumstances.

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The RNA was eluted by adding nuclease-free water into the column and centrifugation of 1 min at full speed. The yielded RNA was checked by nanodrop (Thermo) and the integrity of the RNA was assessed in the Experion automated electrophoresis station

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Finally, the amplified DNA regions are compare using a gel. DNA Profiling

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