Ruby Primer

Polymerase Chain Reaction was run and the results were placed into an electrophoresis gel to visualize the PCR product. To determine the alleles in the gene, a digest enzyme, Fnu4HI, was placed into the DNA. Tasters would view a cleavage formed at a restriction site in at least one of the alleles, whereas non-tasters would not.

Other forms of the genotype, (ygr/ YGR) and (YGR/YGR) will result in green leaves. A third gene in Brassica rapa is the rosette mutant, homozygous recessive. The genotype needed for the short, rosette plant form is (ros/ros). The other two genotypes (ros/ROS) and wild type (ROS/ROS) will result in the normal form of the plant. The phenotypes and genotypes are related in that the phenotypes provide a visible indication of the genotype. This is true in an individual with a homozygous recessive gene. However, in the case of dominant genes, since only one copy is needed for the phenotype to be present, then the second copy is not indicated. The second copy can be identified process where two individuals (P1 and P2) with the same dominant phenotype, called the parental generation, are bred. This produces an F1 or first generation of offspring. The F1 generation can also be bred and produce an F2 generation. Each individual in the F1 and F2 generations receives one copy from each parent of the 3-letter genotype code, called an allele.

Published: September 17, 2002 VeriFone, Inc.® Petroleum/C-Store 300 South Park Place Blvd., Suite 100 Clearwater, FL 33759 Office: (727) 953-4000 Fax: (727) 953-4001 Printed in the United States of America. © 2001 VeriFone, Inc. All rights reserved.

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.

Flow Diagram Homework Assignment Name: Preetam Ganti Using the table below, explain the purpose of each of the steps of the Experimental Flow Diagram (Figure 1 from your lab manual – it is provided on the next page) for answering the two research questions guiding this laboratory investigation: 1. Is the bacterial contamination at these three farms due to the same plasmid or different plasmids? 2. What is the frequency of tetracycline resistant bacteria in the beef farm cultures?

The experiments purpose was to understand and observe the gene expressions in the genes pCNT103, cig1 and GapC in the shoot, root and callus tissues of the tobacco plant, Nicotiana tabacum. Using various genetic laboratory research techniques completed the experiment. The experiment consisted of four parts. The sterile tissue culture technique was used to differentiate the tobacco callus tissue in

In order to determine the type of bacteria we had, be it GST or GST-Cherry, we conducted various experiments through protein and DNA analysis. The purpose of this experiment was to analyze two unknown plasmids (#16 & #23) and, run them through a restriction digest in order run agarose gel electrophoresis as well as a bacterial transformation in order to view bacterial growth. The DNA was also run with BamHI, HindIII, and a mixture of BamHI/HindIII. This resulted in DNA fragment of different lengths from the agarose gel electrophoresis as well as small faint bacterial growth on our agar plates.

To decipher if a species by its morphology can be suggested as a hypothesis, but the results of its DNA will identify the species accurately. Tissues samples can be taken from the species in question, and the DNA can be extracted from tissue. Once the DNA is extracted it can be amplified. DNA can be amplified by the PCR procedure, in which specific gene regions can be used as barcodes to identify the species. These specific regions are known as Cytochrome oxidase 1 and Cytochrome B.

PCR genotyping utilizes restriction fragment length polymorphism (RFLP). This method uses restriction enzymes to cut the DNA at specific sequences. Then, these segments of DNA are amplified through three steps: denaturation, annealing, and elongation. Denaturation breaks the double stand. DNA primers anneal to the DNA template during annealing. During elongation, the DNA primers add nucleotide bases to complement the template strand. Then, place the PCR products int the electrophoresis gel to separate the DNA by size (smaller will move faster, and bigger will move slower). This is the crucial step that will show one if the PCR produced the desired DNA fragment that one is genotyping. The DNA ladder contains the DNA fragments of known size which will be used to compare to the bands produced in the other lanes (the PCR products). If the PCR product contains the DNA fragment, then the PCR bands and the DNA ladder bands should match up. My gel electrophoresis sample did not show due to incorrect techniques. My samples might not have been loaded into the wells correctly. However, one should expect to see the wild-type fly at 467 bp, white fly at 704 bp, and the unknown in lane 3 both the 467 and 704 bps. Female flies are homozygous recessive. As seen in all of the white crosses, female white flies are very rare compared in t number to the male white flies. Therefore, one can infer that female offsprings must inherited 2 alleles for the white eye color to show in females. The WT male x w female cross shows that female white flies inherit the white eye phenotype in a homozygous recessive

XhoI is a type I restriction enzyme, so the location of the recognition site on lambda DNA is unknown but it is known that it cleaves at CTCGAG sites (XhoI (10u/ul)). While the DNA fragment lengths and cleavage sites are known for HindIII lambda DNA digest, they are not known for a XhoI lambda DNA digest. The goal of this study is to determine the recognition site of XhoI on lambda DNA by comparing the DNA fragments from a HindIII digest to the DNA fragments of a HindIII and XhoI digest. We will do this by creating a mixture of solutions containing only DNA, DNA and HindIII, DNA and XhoI, and DNA, HindIII and XhoI. We will then run a gel electrophoresis, which will separate the DNA, fragments by size and we will compare the DNA fragments from each solution. Through comparing the DNA fragments from the HindIII digest and the HindIII and XhoI double digest, we will be able to determine the XhoI recognition site on the lambda DNA.

With the sample wells near the cathode of the electrophoresis chamber, a potential difference of 100 volts was applied across the gel. The chamber was covered with aluminum foil to protect the SYBR SafeTM dye from light. Once the Orange G reached about 80% across the gel, the electrophoresis was stopped and the gel was carefully removed. In order to visualize the results of the PCR and electrophoresis, an image of the gel was taken under ultraviolet light. The resulting bands of the control samples were verified. Using the bands of the control samples as a guide, the presence of the Bt gene was evaluated by comparing the bands produced by the test corn chip.

In Table 1, the data presents the values for calculating the strawberry DNA yield and the DNA yield value. The value of the concentration of the Methylene blue-stained DNA solution is obtained from the standard curve in Figure 1. The value was also obtained using a strawberry DNA extraction process. In this process, a DNA Extraction Buffer containing detergent and salt (sodium chloride) was used. The detergent helped in breaking down the strawberry cells to release the DNA into the solution. Water wasn’t used in this process because the phospholipids in the cell membranes wouldn’t be dissolved. The salt in this process helped in creating an environment for the DNA strands to gather and clump together. This way, it would be easier to observe

The IL-10 promoter polymorphism (PCR-Restriction Fragment Length Polymorphism) assay as described by Turneret al(year) is used for detecting the -592 SNP by introducing 2 alleles, A and C (A-C). A measurement of 100 ng of genomic DNA is amplified in a 25 mL and final volume reaction mixture containing 10X KCL buffer, 1.5mM MgCL2, 0.2mM of each dNTP, 25 pmol of each primer, and 1 unit of Taq polymerase was determined. The cycling conditions were determined as follow: first, in 95°C for 5 minutes; and second 35 cycles of 94°C for 40 seconds at 61°C for 1 minand at 72°C for 40 seconds were specified; finally at 72°C for 10 minutes it was determined. The PCR product contains 412 bp and the Restriction Fragment Length Polymorphism assay was performed in a 15 mL reaction mixture containing PCR product (10 mL), buffer (1.8 mL), enzyme (RsaӀ 2 units per reaction), and distilled water (3 mL). The reaction

Advances in genotyping technology, reduction in cost of next generation sequencing has led to a rapid increase of molecular markers for genomics studies (Bradbury, Zhang, et al., 2007, Chaudhary, Patil, et al., 2015). The use of association analysis to study complex traits in plants is now a viable approach. TASSEL program (Trait Analysis by aSSociations, Evolution and Linkage) (Bradbury, Zhang, et al., 2007) is software used to conduct association analysis. TASSEL accounts for false positives that arise from population and family structure by structured association (Bradbury, Zhang, et al., 2007).

The advantages of DNA based markers are that they are selectively neutral, abundant and have a wide coverage in the genome of all species. DNA based markers have been widely used in genetic studies of various crop plants that include cat’s whiskers (Kafesu, 2012) and tobacco (Mwadzingeni et al., 2013). Several DNA-based techniques can be used for characterization and these include simple sequence repeats (SSR), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD) (Villa et al., 2005). These techniques are all fairly effective. Their choice depends on the stage of research into