Cyp And Gst Polymorphism And Absolute And Relative Quantification Report

A possibility of not getting a fragment under the test food with plant primers slot could be due to the fact of not extracting enough DNA from the sample food. Another reason for any missing fragments could be that the DNA ran off the gel, or that cross-contamination occurred with improper labeling of PCR tubes. These errors could be avoided by easily using the right sanitation methods, following the protocol and ensuring the slurry mix contained a good ratio of sample food to

was used to find the frequencies of the dominant and recessive alleles. After noting and calculating the results of the first fifty people at the mall, I tested a second experiment, deducing that attached ear lobes seemed to be more prevalent in younger females. My new hypothesis was females under age thirty had a higher frequency of attached ear lobes, so I then observed the next fifty females that appeared under the age of thirty and recorded the results. The Hardy Weinberg formula was again used in the calculations.

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 first possible gene is ras1 or raspberry gene. This gene is located on the X chromosome and at 9E1-9E2 on the cytogenetic map. The location on the recombination map is unknown. The ras1 gene is a possible choice due to the dark red and orange color combination that it expresses. (Nash et al. 1994) Another gene is the Pur1 or the purplish ruby gene. This gene is located at 3-39.5 on the recombination map. The Pur1 gene could be responsible for the mutant’s eye coloration due to the red and brown pigments that are present. (Aparisi and Najera 1990) The pr1 or purple gene is another possible explanation for the mutant’s eye color. This gene is located at 38B3-38B3 on the cytogenetic map. It’s location on the recombination map is 2-54.5. The purple gene has a high probability of being responsible for the mutant’s eye color due to the light reddish color combined with a touch of orange and the similar texture that it possesses. (Kim et al. 1996) Another possible gene is ry1 or rosy. This gene is located on the 3R chromosome. Its location on the recombination map is 3-52.0. The rosy gene can be located at 87D9-87D9 on the cytogenetic map. Rosy gene is another possibility because it possess a red shade with more orange and brown pigments that could compensate for the lighting when viewing the mutant specimen through the stereoscope. (Zhou and Riddiford

The site of crossing over occurred most often on chromosomes a, b, c, and d, while crossing over occurred less often on chromosomes e, f, and g. This is shown by the data, because chromosomes a, b, c, and d had the highest numbers and percentages of being on the same chromosome as “a”, while the opposite is shown for chromosomes e, f, and g. The lab results also showed that the recessive allele “a” appeared most often with “a” because in order for any crossing over to occur, the recessive allele “a” must be present. This is shown in the die because the die is guaranteed to roll at least a 1.

Lane 3 contained the initial PCR product, which was not successful. Lane 4 showed the nested PCR, which was successful and resulted in three distinct bands. The accompanying table (table 1) showed what was loaded into each lane, the number of bands, and the approximate base pairs (bp) present in relation to the molecular ruler. The gel was loaded with samples from the entire lab table. Lane 1 and 2 showed Lydia's results for initial and nested PCR on the same plant, but from a different original

The goal of this experiment was achieved because the subject’s genotype for PTC, with a phenotype of taster, was determined using PCR and gel electrophoresis. The genotype was heterozygous dominant (Tt) based on the number of DNA fragments in Figure 1. Therefore, the gel picture matched the prediction and hypothesis that either three or two fragments would be present since the subject was a taster for PTC. The DNA fragment amplified by PCR was about 300 base pairs long, 303 bp to be specific. The enzyme, Fnu4HI, cut the DNA at a restriction site 5’-GCNGC-3.’ Tasters would have a C in the amplified fragment, while non-tasters would have a T. Once the DNA was cut, the fragments created were 200 and or 100 base pairs long if the DNA was from a taster. This is what occurred for the subject as seen in

Always use two letters to represent a genotype. There are two letters in a genotype because one gene comes from the mother and the other one comes from the father. The three possible genotypes are two capital letters(TT), one of each(Tt), and two lowercase letters(tt). Homozygous means two capital letters, or two lowercase letters. A homozygous genotype is (TT), or (tt). Homozygous, also means purebred. Heterozygous means one capital letter and one lowercase letter. A heterozygous genotype is (Tt). Heterozygous, also means hybrid. (TT)homozygous=purebred, (Tt)heterozygous=hybrid, (tt)homozygous=purebred. How it physically shows up is a phenotype. Examples of phenotypes are blue eyes, brown fur, striped fruit, yellow flowers. Controlled by two or more genes is polygenetic inheritance, it may be on the same or on different chromosomes. Three examples of this, are eye color, skin color, and blood group. They are located in corresponding positions on homologous chromosomes called loci. A dominant allele hides the effect of another allele. We represent this, with a capital letter. A recessive allele is hidden, unless it is placed with another

I predicted that these two populations would be opposite in the alleles they carry. The Greek population would be majority homozygous recessive (no insertion), and the Taiwanese population would be majority homozygous dominant (Alu insertion). The pictures of these tests are shown

For -629C/A polymorphism, 5µl of PCR products were incubated with 0.4U of Van91I restriction enzyme at 37 °C for 48 h. The thermocycler conditions after optimizing the technique for -629 C/A were: initial denaturation at 96°C for 5 min followed by 35 cycles of amplification, each cycle consisting of 45 s at 96 °C, 45s at 58 °C and 45 s at 72 °C, in a PTC- 200 MJ-Research Peltier thermocycler. The thermocycler conditions after optimizing the technique for Taq1B were: initial denaturation at 96 °C for 5 min followed by 35 cycles of amplification, each cycle consisting of 45 s at 96 °C, 45s at 56 °C and 45 s at 72 °C, in a PTC- 200 MJ-Research Peltier thermocycler. The reactions for both polymorphisms ended with an additional 7 min of extension at 72° C. The Taq1b PCR resulted in an amplified product of 535 bp and was digested using the restriction enzyme Taq1B. The presence of the Taq1B restriction site produces the B1 allele pattern

This relates to this experiment in that the white-eyed sex-linked trait is the recessive gene being traced back to the parent generation. The objective is to identify the parent generation’s genotype from only knowing the F1 generation’s phenotype and conducting a cross between sixteen of the F1 generation to produce an F2 generation. The F2 generation should be able to solve the P0’s phenotype. It should also be supported by statistics, punnett squares, and a low probability value. Based on the phenotypes in both F1 and F2, the parent generation’s genotype should be homozygous white-eyed females and dominate red-eyed males. Methods

Extracting DNA from available cells is a widely used practice with applications such as forensics, genetic testing and victim identification to name a few. DNA extractions have been widely practiced for decades. This report simply summarises our results comparing two methods.

Random amplified polymorphic DNA (RAPD) markers are analyzed by using PCR to amplify the segments of nuclear DNA. The use of a single primer (usually 8–10 bp long) that attaches to both strands of DNA and low annealing temperatures increases the likeli¬hood of amplifying multiple regions representing a particular locus (Welsh and McClelland, 1990). Although RAPD is a simple and inexpensive technique its major limitation is the inability to differentiate between homozygote and heterozygote; this marker is therefore regarded as a dominant type. This technique is based on the use of short, arbitrary primers in PCR reaction and can be used to produce relatively detailed and complex DNA profiles

PCR tubes were ready for purification when a single DNA band was present per lane on the agarose gel. If multiple bands were obtained in a single lane the PCR process and gel visualization process was repeated however, the denaturation temperature was run at a higher temperature to prevent non-specific binding of the primers to the DNA template. The purification process used an Invitrogen PCR clean-up kit. 5mL of charge switch beads

in exon 5, *4 (1846G>A in intron 3 - exon 4 junction), *5 (whole gene deletion), *10 in exon 1(188C>T, P34S), *14 (1758G>A), *17 (1023C>T; 2850C>T) in exon 2 and *41 (2988G>A) variants were the most characterized alleles of CYP2D6 (http://www.cypalleles.ki.se/) consists of three genes: (1) CYP2D6, a “functional” gene; (2) CYP2D8, a pseudogene that arose by gene conversion; and (3) CYP2D7, a nonfunctional gene that arose by gene duplication (Figure 13). In