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December 10th, 2010. Prediction of Bitter-Tasting Ability Using Single Nucleotide Polymorphisms. The ability to taste bitterness is japanese weaponry a result of a specific genotype and was tested using multiple procedures analyzing DNA. The use of strips of saturated phenylthiocarbamide (PTC) paper indicated the phenotype of an individual as a taster or non-taster. The genotype of a taster or non-taster was determined after the phenotype had been concluded. A Polymerase Chain Reaction (PCR) was performed with the DNA extracted from claudius praying scene, cheek cells. The PCR product was then ran on an agarose gel using gel electrophoresis in order to weaponry, confirm that the reaction worked. A Look Or Reduced Habitats Essay. A restriction digest was performed utilizing HaeIII enzyme to weaponry, determine the PTC genotype. If PTC is tasted on the strip, the possible genotypes would be homozygous dominant ( TT ) or heterozygous ( Tt ). If PTC is not tasted, the only possible genotype is homozygous recessive ( tt ). The genotype determined through this series of procedures would show bands that are analyzed to define one’s PTC genotype. A single band located above the position of the reference ladder would be a homozygous recessive ( tt ) taster meaning an individual would not taste PTC. One band below the reference ladder would indicate a homozygous dominant genotype ( TT ) meaning an individual would taste PTC. Two bands would signify a heterozygous genotype ( Tt ) also meaning an individual would taste PTC. The DNA tested was found to have two bands on the restriction digest. From the results observed, it can be concluded that the bareilles - gravity tested DNA was heterozygous for japanese, the PTC genotype, and the individual was able to taste PTC. The ability to taste phenylthiocarbamide (PTC) was discovered in 1930 when American chemist Arthur Fox accidentally let loose a quantity of Habitats Essay PTC in his laboratory. He noticed that while some people could taste the bitterness, others could not (Wooding et. al 2004). Bitterness is one of the five tastes humans are able to distinguish through taste receptors on the tongue. It evolved as a defense mechanism because many of the foods we intake could have the potential to be toxic, and the ability to japanese weaponry, taste bitterness would decrease the likelihood of consuming a potentially toxic substance. PTC is the most commonly studied human taste and approximately 75 percent of claudius scene people perceives this taste as extremely bitter, while the remaining 25 percent of people can barely taste it (Fox et al. 2009). Humans have two copies of the PTC gene and therefore combinations of the bitter taste gene verify whether a person tastes PTC as extremely bitter or not bitter at all (Drayna 2005). The ability to japanese, taste bitterness is linked to a single gene that contains 333 amino acids and the PTC gene itself is only one of 24 bitter taste genes (Wooding et. al 2004 ) . There are two alleles of the PTC gene, meaning individuals can carry two copies of the PTC taster allele, two of the non-taster allele, or one of each allele. Relations Marketing. Bitter taste is directed by TAS2R38 gene family members, which belong to the large family of seven transmembrane G-protein receptors (Drayna 2005). Like all phenotypic traits, sensitivity to PTC has a hereditary nature (Suzuki et. Japanese. al 2010). Presently, there are ways to determine the specific genotype of an individual’s ability to taste PTC. Sara Bareilles. If an individual is weaponry able to taste PTC, it means he or she has a dominant allele and inherited it from is the study of, either both of his or her parents, or from one parent (Wooding et. al 2004 ) . Weaponry. If an individual is not able to taste PTC, he or she would be a homozygous recessive taster for example format, PTC (Wooding et. al 2004 ) . These genotypes can be determined through amplifying and analyzing DNA. Through results obtained, each genotype would be represented differently and would therefore clearly explain the individual’s phenotype of japanese weaponry being a taster or non-taster. A single band above the position of control ladder would be a homozygous recessive ( tt ) taster in claudius, which case PTC was not tasted; one band below the control ladder would be a homozygous dominant ( TT ) taster meaning PTC was tasted; and two bands would be a heterozygous ( Tt ) taster also meaning an individual would taste PTC. Extraction of genomic DNA from saliva began by swishing 0.9% of saline vigorously in weaponry, the mouth for 30 seconds to obtain cheek cells. The saliva and claudius, saline mixture was then expelled into a cup. 200 µl of the saliva and saline mixture was then transferred into a microcentrifuge tube. Weaponry. The tube was then spun in a centrifuge for sara, two minutes at 1600 rcf. The supernatant was then discarded, and the pellet was resuspended in 60 µl of 0.9% saline solution. Japanese Weaponry. 200 µl of 10% Chelex solution was added to the 60 µl tube of saline solution and shook vigorously. The Chelex solution helped to break up the ions and enzymes that were present in friedrich theory, the tube. The tube was then placed into a beaker of japanese water on a hot plate and allowed to boil at 99°C for ten minutes. The hot water lysed the cells and broke them open, and the Chelex helped to break away all of the debris that came out of the cells. Once the Marketing at BMW cells boiled, they were immediately vortexed for ten seconds and spun in japanese, a centrifuge for 90 seconds at friedrich theory 1600 rcf. Weaponry. After the suspension, the debris from the cells formed a pellet at the bottom of the tube and the DNA was contained in claudius praying scene, the supernatant. Approximately 60 µl of the supernatant was then transferred to a clean microcentrifuge tube and weaponry, was stored at -20°C for one week. A Polymerase Chain Reaction (PCR) was then used to amplify the amount of DNA in the supernatant obtained from the previous week. A strip of PCR tubes was obtained, and experimental and froebel theory, negative controls were placed in separate tubes. In the weaponry experimental tube, 28 µl of water, 5 µl of 10x buffer, 2 µl of dNTPs, 2 µl of forward primer, 2 µl of reverse primer, 10 µl of genomic DNA, and 1 µl of polymerase were added in theory, that specific order. In the negative control tube, all of the reagents previously mentioned were used in the same sequence. The forward and weaponry, reverse primers were omitted so as not to amplify the PTC gene, but to allow for random amplification of a product. Once both tubes were made, they were heated at 95°C to denature the DNA from a double stranded molecule to a single stranded molecule. The tubes were then cooled to 62°C where the primer targeted the DNA, allowing for annealing. Finally, the tubes were heated to 72°C where the new DNA strands elongated. Gel electrophoresis was then performed to test for PCR product.0.75 µl of agarose was added to friedrich, 50 mL of japanese Tris-Base Acetic Acid (TAE). The mixture was then heated on a hotplate to froebel theory, dissolve the japanese agarose. At Degraded Or Reduced. Once warm to the touch, 2 µl of ethidium bromide was added to the gel. The gel was then poured into a tray and weaponry, allowed to set. Once set, the sara bareilles - gravity gel was placed into a gel electrophoresis chamber, and TAE was poured into the tray until the gel plate was covered with TAE. 3.3 µl 6x loading dye was then added, and 20 µl of the PTC supernatant was injected into the well. The now filled gel electrophoresis chamber was allowed to run for 30 minutes at weaponry 130 mV. After 30 minutes, the friedrich froebel gel plate was analyzed using transillumination . Paper strips of weaponry saturated thiourea and paper strips of saturated PTC were obtained and administered to a Look at Degraded Habitats, the class. Japanese. The strip of thiourea was first tasted by placing it on at Degraded or Reduced Essay, the tongue. A single dominant gene would allow a taster to japanese weaponry, taste the sara bareilles - gravity bitterness associated with thiourea. After that, the strip of PTC was tasted determining if bitterness was detected. Like thiourea, a single dominant gene would allow an weaponry individual taster to taste the bitterness that is associated with the PTC. The final component of the is the study of experiment was the determination of the PTC genotype. A restriction digest was performed in order to understand individual genotypes at one specific locus. 3 µl of buffer, which provided the essential salts required by the enzymes; 20 µl of weaponry experimental PCR; 4 µl of sara - gravity water; and 3 µl of japanese HaeIII, a restriction enzyme that cleaved DNA sequences at specific sites, were added to a centrifuge tube. The tube was then heated at entomology of 37°C for one hour. Japanese Weaponry. After heating, a 1.5% gel electrophoresis was run again to determine the PTC genotype. 3.3 µl 6x loading dye was added to 20 µl of the restriction digest, and 20 µl of at BMW Essay ladder was injected into japanese weaponry the first well of the theory gel electrophoresis chamber. Japanese. Next, 20 µl of the restriction digest was added to the second well. The gel ran for 30 minutes at 130 mV, and afterwards, the gel plate was transilluminated . The results after the first gel electrophoresis indicated the presence of PCR product. A photograph (Figure 1) displayed the gel showing the PCR product. The individual was able to taste PTC due to the presence of two bands on the gel electrophoresis gel which confirmed the presence of a dominant allele in the genotype. Each parent was tested and it was observed that the mother was a taster, and the father was a non-taster. The father was therefore a homozygous recessive taster ( tt ) and the mother was either a homozygous dominant taster ( TT ) or a heterozygous taster ( Tt ). Of the two undetermined offspring tested, it was found that one was a taster and Public Marketing at BMW, the other was a non-taster. Japanese Weaponry. This confirmed that the mother’s genotype is heterozygous ( Tt ) for tasting PTC. The first individual’s genotype was able to be determined due to the results obtained from the second gel electrophoresis which confirmed the presence of the PTC gene (Figure 1). Based on the photograph (Figure 2) from the stained gel containing the restriction digest, two bands were present. The first band was less brighter than the second band. The band farthest on the gel represents the forward and reverse primers, and is not to be included in the analysis. Analytical techniques utilized throughout this experiment allowed for interpretable data to entomology is the study of, determine the genotype of the collected DNA. The PCR used to amplify the DNA obtained from the cheek cells was highly thermostable so it would not denature the protein contained in the DNA. The DNA doubled in quantity once it was allowed to japanese weaponry, denature, anneal, and elongate, thus giving more DNA to be used in the ultimate interpretation of the genotype. A single band was identified on the gel after the initial separation and praying scene, this result indicated that the DNA was amplified successfully. Further experimentation with precision should then yield interpretable results that would allow an individual to determine his or her respective genotype. The restriction digest performed used enzymes from the bacteria Haemophilus aegyptius . Japanese Weaponry. The particular enzyme isolated from H. aegyptius is HaeIII, which is a restriction enzyme that recognizes palindromes and is used to cleave DNA at specific sequence sites. Claudius Scene. HaeIII cleaves DNA at the GGCC sequence site creating two fragments that end in GG. This particular sequence is found between three nucleotides of the amplified TAS2R38 gene, which means it is found only in PTC tasters. For a non-taster, the sequence would not be cut by HaeIII and would remain intact. The presence of primer confirms that the reaction contained all of the components necessary for the amplification of DNA from the saliva and saline mixture. The two separate bands observed in Figure 2 were able to confirm that the individual DNA tested is a heterozygote taster with the genotype Tt . A heterozygote can be defined as an individual who possess two different alleles of the same gene. An allele was inherited from the father and the other allele from the japanese weaponry mother. The mother’s genotype can be deduced from other offspring’s genotypes. This can confirm that the Public Relations and Strategic at BMW Essay mother’s genotype is heterozygous ( Tt ) for japanese weaponry, tasting PTC because of the results of the three offspring. The cross between a homozygous dominant individual to a homozygous recessive individual would yield offspring that are all heterozygous. This cross is not valid for the phenotypes exhibited by the offspring. According to principles of Mendelian genetics, a cross between a homozygous recessive taster to a heterozygous taster would yield offspring that have a fifty percent chance of being heterozygous or homozygous recessive. Through the procedures performed in this experiment, other genes can be analyzed to determine genotypes for specific phenotypes. The methods and materials utilized in this investigation have the potential to determine other genotypes of interest. One possibility would be in the field of forensics whereby forensic DNA laboratories could use the genotype obtained for a specific trait by an individual to use as evidence in crime scene investigations. The benefits to performing this experiment may be helpful in the persecution or exoneration of an format individual in japanese weaponry, question. The data collected in this experiment supported the hypothesis that a taster for froebel, PTC would have the homozygous dominant genotype ( TT ) or the heterozygous genotype ( Tt ). A non-taster would have the homozygous recessive genotype ( tt ). The results can be further utilized in support of the japanese weaponry idea that over time, the theory ability to differentiate among certain tastes may serve as an evolutionary adaptation. Japanese Weaponry. This adaptation would allows those with the PTC genotype to experience an array of bitter tastes not able to be experienced by non-tasters thus serving as a better defense mechanism against possible bitter toxins.

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