The second step performed was the Gram Stain test. The Gram Stain was used to determine whether the unknown bacterium was gram-positive or gram-negative. First, the unknown sample was smeared onto a slide along with a drop of distilled water. Second, the unknown was air dried and was heat fixed.
Amino acids and proteins are more negatively charged at equilibrium than in stacking gel. As a result,
Transformation in bacteria usually takes place when a bacterial cell accepts strange DNA and integrates to its own DNA. The transformation normally takes place within plasmids, which are tiny circular DNA molecules that have been separate from its own chromosome. The copies of the same plasmid range from 10 to 200 copies within a cell. These copies of plasmids may multiply when the chromosome replicate or multiply independently. One plasmid has a range of 1,000 to 200,000 base pairs.
The objective of this experiment was to determine the identity of three unknowns existing in a 1:1:1 mixture. An extraction technique was therefore used to identify the unknown acid, base, and neutral as the compounds must first be separated. Extraction is a method of separation that is based on the difference in solubility of each compound in a mixture. There are a couple of necessary requirements for an extraction to be completed with experimental success. In this extraction there were two liquid solvents used, an original solvent (So), and an extraction solvent (Sx).
The purpose of this lab is to determine the relationship that exists between the number of amylase gene copies and ancestral diet. As the human civilization moved forward toward agriculture the diets of humans also changed. Depending on where the humans originated would give insight to how much of their diet was starch based. My family’s geographic origins are from China. Thus knowing that the country has a high starch based diet, we would suggest that I would have a high amylase production.
Before Gel Electrophoresis, separation of small molecules was impossible. Today Gel Electrophoresis is the primary method of separating molecules. The ability to separate has greatly improved forensics. paternity/maternity tests, and many other useful tests. Prosecutors being able to prove that a crime was committed because of DNA instead of testimony has improved the criminal justice system greatly.
Once the loading solutions were made and the agarose gel electrophoresis apparatus was set up, the solutions were dispensed into the wells of the gel, as well as a reference solution. The gel image gathered from the gel electrophoresis allowed for the analysis of the plasmid DNA and the effects the restriction endonucleases, Hind III and Hinc II, had on it. While analyzing the gel image, it was apparent that based on the reference solution in lane 1 and the mark in lane 3, the Hind III solution, that the number of base pairs should be the same. Also it was apparent that lane 5, the Hinc II solution, had 4 marks. By measuring the migration distance in centimeters and plotting against the log(# of base pairs(bp)) for the standard in lane 1 the linear trendline equation provided by the calibration curve allows for the number of base pairs to be found for the marks in lane 3 (Hind III) and lane 5 (Hinc II).
Abstract: Gel electrophoresis is a method used to separate DNA fragments according to size. During our study we pondered on one particular question; whose blood was left at the crime scene in the AP biology classroom? Before carrying out our experiment we learned about the process of gel electrophoresis and the use restriction enzymes. After analyzing our results, we decided to reject our hypothesis because our experiment showed strong evidence against what we originally hypothesized.
Introduction. Christian Gram created the Gram stain technique in 1884 (K. Rudolph [Clemson University, Clemson, SC], personal communication). Gram used tissue in his first attempt with staining (1). The staining of the organisms allowed for the viewing of structure and determining those components by the viewing of the color that appeared after staining the organism (2). The purpose of this experiment was to identify the organism as gram-positive or gram-negative.
Materials & Methods To determine the presence of tissue plasminogen activator (TPA) regions on chromosome 8, we prepared a polymerase chain reaction (PCR) to amplify our DNA samples and used gel electrophoresis to visualize the results. Samples of cheek epithelial cells collected by rinsing our mouths with 10 mL of a 0.9% NaCl solution for 30 seconds were used as the template DNA for the reaction. Using a 100-1000 μL pipettor, two increments of 750 μL of the expelled salt and cheek mixture were transferred into a labelled 1.5 mL microfuge tube. Tubes were collected and centrifuged at 13 000 rpm for 10 minutes. Following centrifugation, the supernatant in the tube was discharged into the sink and the tube was placed in an ice bath.
A0123942_Gel Electrophoresis Report Name: Lee Zixuan Process of Gel Electophoresis: Gel electrophoresis in this case involves the placement of both genomic and plasmid DNA inside the wells of the agarose gel, together with a gel loading buffer. The agarose gel contains mini pores such that when an electric current is switched on, it would be able to separate the bigger segments of DNA bands from the smaller ones. As DNA is negatively charged due to the phosphate group, it would move towards the positive electrode. Smaller molecules of DNA would be able to move faster than the bigger molecules of DNA. Upon completion, the separated fragments of DNA can be visualized under UV light and the distinct bands could be seen.
Experiment #7: Column Chromatography of Food Dye Arianne Jan D. Tuozo Mr. Carlos Edward B. Santos October 12, 2015 Abstract Column chromatography is the separation of mixture’s components through a column. Before proceeding with the column chromatography itself, a proper solvent system must be chosen among the different solvents. The green colored food dye is the mixture whose components are separated.
Abstract This experiment was carried out to determine the species of the unknown organism. Once a choice of the unknown was made a Gram stain was conducted to determine the gram nature and morphology of the organism which was Gram negative bacilli. Based on those results a citrate utilization test was performed resulting in a positive test. Following the flow chart the next test to conduct was a motility test which also had a positive outcome.
The purpose of this lab was to synthesize 2-nitrophenol and 4-nitrophenol from phenol, sodium nitrate, and sodium nitrite, as well as to use column chromatography to isolate the two products from one another and the byproduct 1,4-benzoquinone. The reaction was monitored by Thin Layer Chromatography to track the progress if the refluxed reaction. Column chromatography was used as a very effective technique for separation as both solids and liquids can be separated by column chromatography. The two nitro phenol products were compared using melting point determinations and the products were also classified and checked for impurities by determination of IR and 1H NMR spectra. Results and Discussion Column chromatography is the most useful purification
A direct current is passed through the gel, and it passes through electrodes at each end of the chamber. Since DNA is negatively charged, it will be attracted to the positive pole and repelled by the negative pole. Small DNA molecules are able to move through the gel faster than larger fragments. This results in a series of
