Phenotype Of Taster Essay

For millions of years, repetitive DNA sequences such as the “Alu sequence” have been randomly inserted in the human genome. In this experiment, I will study whether or not the repetitive element such as the “Alu sequence” is present in the PV92 region of chromosome 16 of my DNA. I will then gather the data from my Biology Lab Class and compare it with those of the United States. To perform this experiment, the method of Polymerase Chain Reaction (PCR) will be used to amplify the DNA. The purpose of using PCR is to create enough mass in order for it to run on the agarose gel electrophoresis. Performing this

The following results helped obtain the haplogroup that in which the sequence of mtDNA would identify. The PCR reaction worked, and this can be determined by looking at the agarose gel in figure 1. If the PCR reaction was successful, than a band should appear around 550bp. Individual AC displays a band around 550bp, this means the PCR reaction was successful. The band for individual AC, depicts a low concentration of product, because the band faint. After the purification process the concentration, A260/280 ratio, and A260/A230 ratio were determined by using the nanodrop. The concentration of mtDNA in the product was 60.9 ng/uL. The ratio for A260/280 was 1.79 and the ratio for A260/230 was 0.77. The A260 and 280 are a spectrometer measurement that measure absorbance at wavelengths of

Four microcentrifuge tubes were placed in a rack, labeled and numbered, in order to identify the group and the DNA/restriction enzyme that it held. Each of the tubes initially received 10 microliters of reaction buffer. There were two samples of suspect DNA provided along with two restriction enzymes (EcoRI and HindIII). Tubes labeled 1 and 2 received 15 μL of DNA from suspect one while tubes 3 and 4 received 15 μL of DNA from suspect two. Following that, 15 μL of Enzyme 1 (EcoRI) were added to tubes 1 and 3, and 15 μL of Enzyme 2 (HindIII) were added to tubes 2 and 4. (Table 1). The tubes were then gently tapped on the counter to mix the DNA and enzyme solution followed by incubation at 37°C for 45 minutes. After incubation, 5 μL of 10x gel loading dye were added to each of the four tubes of suspect DNA. The tubes were then placed on ice while the gel was under preparation.

There were several steps used to acquire the colony necessary for the PCR. First a student forearm was swabbed using a cotton swab, the cells were then placed in an agar plate. DNA was then extracted from the cultured bacteria by using a technique to lyse the cells and solubilize the DNA, then enzymes were used to remove contaminating proteins. The DNA extraction consisted of a lysis buffer that contained high concentrations of salt for denaturing. Binding with the use of ethanol and a washing step to purify the DNA. The final step for the DNA extraction was elution where the pure DNA was release. Proceeding the extraction of DNA the results of the 16s gene amplification were examined through gel electrophoresis it was analyzed by estimating the size of the PCR bands with marker bands. After measuring the success of the extraction, a technique called TA cloning was started. Cloning of PCR products was done by using partially purified amplified products with

The DNA extraction results, along with the PCR product, did not fare well. There was not enough product produced to be viable in the later stages of the experiment, so a backup was used in place of the original product.

When picking up the gel, the gel broke in half. This could have altered the results because some of our DNA could have broken off. Although it appeared that we saved the DNA, there is a slight chance that some of the DNA could have broken off. Also, my assistants and I had difficulty measuring the agarose powder. We might have poured too much of the powder, which could have made the gel too thick. Next time, we will be more cautious when measuring the powder to have more accurate results.

Phenylthiocarbamide (PTC) is a substance that can only be detected in human tasting receptors if the individual has a dominant allele for that trait. This experiment analyzes a small sample of college students and uses PTC taste testing to assess the number of strong, moderate and non-tasters for PTC. It uses polymerase chain reaction (PCR) to amplify these students’ DNA strands and assess their genotypes for the PTC tasting gene. The results show that strong tasters seem to exhibit a homozygous dominant genotype for the PTC tasting gene, while moderate tasters exhibit a heterozygous genotype, and non-tasters exhibit a homozygous recessive genotype for the PTC tasting train. The frequency of these alleles are predicted using the Hardy-Weinberg

They were run at 1X 94ºC for 3 minutes, 30X at 94ºC for 30 seconds; 50ºC for 30 seconds; 72ºC for 45 second and 1X at 72ºC for 5 minutes. The PCR reactions took about 1 hour and 30 minutes to complete. The PCR products, were then purified by removing the leftover primers, nucleotides and salts. 250 µl of Buffer BB were added to Tube B and the mixture was pipetted into a spin column. The mixture was centrifuged for 30 seconds at room temperature. Then 2 cycles were completed at 30 seconds each with 200 µl of Buffer WB to remove any impurities. Then 25µl of Buffer EB were added to the tube to release the pure DNA and the mixture was centrifuged for 30 seconds. As the PCR reaction was running, a microscope slide was prepared from the live bacterial culture to observe the individual cells of the unknown bacteria and determine its

Nonetheless, these changes may occur in the regions that regulate these genes and have an effect on their functions, thus allowing multiple alleles to be formed. There are three common SNPs that are correlated to PTC sensitivity, and each one of them causes a change to the amino acid sequences of the PTC receptor (Sullivan, n.d). These SNPs in terms of base pair positions are 145, 785 and 886, and the subsequent SNP-containing codons are proline or alanine; alanine or valine or valine and isoleucine. The SNP haplotypes for the PTC gene include PAV (taster), AVI (non-taster) and

Blood samples were obtained from all family members. Genomic DNA was extracted from 100-μl samples of peripheral blood mononuclear cells incubated with proteinase K in lysis buffer, followed by phenol/chloroform extraction and ethanol precipitation. Finally, the pellet was dissolved in 30 μl sterile water and stored at -20°C until further tested. To determine mutation, specific primer sets within the APC gene were designed (primer sequences upon request). Two microliters of DNA were used to set up 25-μL PCR reactions by using the Perfect Taq plus MasterMix (5 prime GmbH, Hamburg, Germany). The amplification reaction was carried out under the following conditions; pre-incubation at 95°C for 3 min, followed by 35 cycles comprising denaturation at 95°C for 30s, primer annealing at 55°C for 30s, and extension at 72°C for 1min, and concluded by a final extension at 72°C for 5 min. Upon electrophoresis in 2.0% agarose gel stained with EtBr, the amplification products were visualized under UV

In order to test whether the curry leaf possesses one of the 6 alleles and whether it has a Ruby gene promoter, PCR and gel electrophoresis was used. Primers, short DNA sequences designed specifically for each allele of the Ruby gene, were used to isolate the gene and amplify it for PCR. Multiple PCRs were performed, each using a different primer set. Primers attach to specific regions surrounding the gene and amplify only the region that is flanked. The forward primer is bound to a specific sequence before the Ruby promoter, found to precede the actual gene sequence, and the reverse primer is bound to the DNA after the gene. In order for visible bands for gel electrophoresis to be produced, a forward primer must bind to one strand of DNA and a reverse primer must bind to the strand complementary to the one the forward primer binds to. If both primers bind to the same strand, the PCR product formed will not produce a result after gel electrophoresis. The size of the gel band of the DNA amplified by the primer will determine the identity of the allele. Band sizes will

Also (5μl) of DNA template that extracted from stool samles was added then 1.5 μl of each type of Primers(forward and reverse)added to the master mix and then blend well using Exispin vortex centrifuge ,then this tubes would transferred to the Thermocycler machine, which has been programmed by the previous program for amplified of ITS1 region.The PCR products were electrophoresed in agarose gel and visualized on UV trans illuminator and then photographed using photo documentation .

I added 1 g of the agarose powder to 100 ml of TAE buffer in an Erlenmeyer flask. Using a magnetic stir bar, the agarose is heated until it is a completely clear solution. The agarose is poured into the chamber to make wells deep enough to hold 30 µl of solution. I inserted the comb in the chamber and waited for the gel to harden. To prepare for the samples, 10 µl of the XC loading buffer was added into the PCR tube. The professor demonstrated the techniques to load the gel with the DNA samples. After learning the techniques, I added my PCR sample in one the the wells. To run the gel, the power supply was set to 100V for approximately 30 minutes. To stain the gel, the gel was removed from the electrophoresis chamber and was placed into the FastBlast staining tray. The gel was placed in the staining tray for 2 minutes. Then the gel was rinsed for 10 seconds in Wash Tray 1. After rinsing for 10 seconds, the gel was washed for 5 minutes in Wash Tray 2. After agitating the gel for every minute in Wash Tray 2, the gel was washed for another 5 minutes in Wash Tray

This allowed us to sequence an informative region of the TAS2R38. Restriction fragment length polymorphisms were created using the restriction enzyme HaeIII. Gel electrophoresis of the amplified and cut DNA segments provided us with a genotype associated with each phenotype.

PCR reaction was carried out in a 25 μl reaction mixture under standard condition, contained 1x buffer, 0.2 mM of four dNTPS, 1.5mM mgcl2, 1 pM of forward (sequence) and reverse (sequence) primers and 50ng of genomic DNA. Initial denaturation at 95˚C for 4 min was followed by denaturing at 94˚C for 30 seconds, annealing at 60˚C for 30 seconds and extension at 72˚C for 30 seconds for 32 cycles. The final extension was 8 min. The amplicons were run on an ABI 3130x (Applied Biosystems, Foster City, California,