Polymerase chain reaction

process. We then added primer to the pcr tube. We also added nucleotides. The last thing we added to the pcr tube was DNA polymerase. This reads the DNA code. DNA polymerase is selected as it can withstand the heat of the pcr reaction. The pcr tube is then put into a thermal cycler. This can accurately heat and cool the tube which having the right temperature is crucial for the reaction to work. The first cycle begins, this is when the double helix separates, creating two single strands of DNA.…

Introduction Studying the phylogeny of Mesoamerican spider monkeys (Ateles geoffroyi), in relation to other spider monkeys, has been controversial because studies have failed to agree on what characteristics make these species separate or have failed to present complete taxonomic data. Additionally, researchers found little agreement between the sequence data and the previous taxonomic data, to infer phylogenetic relationships. The researchers who performed this study used mitochondrial DNA…

Multiple Myeloma (MM) is characterised by malignancy of antibody-secreting B-cells that accumulate in the bone marrow . Tumour growth promotes hypoxia in the tumour environment. Hypoxic inducible factors (HIFS) are heterodimeric proteins composed of an inducible alpha-subunit and a constitutively expressed beta-subunit. In this experiment, the hypothesis being tested was if HIF1α and HIF2α regulate different genes in multiple myeloma. This was tested using the cross chromatin immunoprecipitation…

DNA fingerprinting, also known as DNA profiling, is identifying an individual using there DNA. This is often used for identifying criminals, and parental testing. First, a sample must be provided, which can be: blood, semen, hair roots, or saliva. The cells from the sample are then split open, and DNA is separated from the rest of the cell. The DNA is then treated with specialized proteins (restriction enzymes),which separate the DNA into smaller parts. The fragments of DNA are then applied to a…

Agarose gel electrophoresis of DNA is used to seperate mixtures of protein and nucleic acid fragments. Molecules tend to differ in size, charge, and mobility. The objective of this procedure is to predict the distance of DNA migrating towards the positive electrode in the electrophoresis chamber. Many scientists use agarose gel electrophoresis to examine the DNA of criminal suspects. The first step in this procedure is to prepare a buffer containing 1% of the liquid agarose gel. Once the gel…

Upon completion of the PCR, the final product should be highly concentrated DNA corresponding to the gene of interest. We ran our gene of interest on gel electrophoresis to compare our gene of interest to a marker of known size to confirm the size of our genomic DNA. On the image of the gel (figure 1), the sample containing the DNA ran, but the sample containing the DNA ladder did not. As a result, an appropriate determination of the sizes cannot be made because there is no ladder to compare it…

RESULTS Transformation: the transformation of the ade2 gene to the kanamycin resistant gene (ade2::kan2R) cause the cell to become red and grow on mediums containing G418 or kanamycin resistant mediums was observed to have occurred. PCR and Gel Electrophoresis: Figure 1 is the product of gel electrophoresis containing the wild type and transformed PCR products, either being or not being cut by HindIII. From the left (reader’s left) of the gel to the right the lanes are; 1.Ladder, 2. transformed…

2.5. Morphological Identification/Evaluations of Entomopathogenic Fungi The six isolates of fungal entomopathogens were provided by Department of Entomology, Penn State USA. Preliminary morphological identifications of fungal entomopathogens such as color of colony, mycelial characteristics and spores from all six isolates were done at microscopic level. 2.6. Molecular Characterizations of Entomopathogenic Fungi Cultures of Metarhizium fungus were grown in nutrient broth (Bacto, Australia) in…

A DNA ladder is a solution of DNA molecules of different but known length. In Well 1, the DNA ladder was applied to the gel to set a standard for the rest of the samples. In LoggerPro, after setting an origin, the second step was to set a standard ladder. We standardized the ladder by going from 4 to 5 cm and determining that was 10 milliliters. The next step was to scale the ladder by selecting the 3 brightest points and labeling it 3000,1000,500 in descending order. The estimated size of my…

Materials 1. Micropipette 2. Micropipette Tips 3. WARDS’S quick view DNA Stain 4. Prepared Agarose 0.8% 5. Tris-Borate-EDTA 5x Buffer 6. Electrophoresis Mold 7. Electrophoresis comb 8. Electrophoresis chamber 9. Electrophoresis power pack and cords 10. DNA from cow liver 11. DNA from chicken liver 12. 224 grams of fresh cow’s liver 13. 224 grams of fresh chicken liver 14. Meat tenderizer 15. Kitchen knife 16. Blender 17. 2.5 ml Salt 18. 31 ml dishwashing liquid (Dawn) 19. 354 ml warm water 20.…