Unknown mixed microorganisms were collected from a building sample, air and fingerprints. The sample microorganisms were swabbed onto petri dishes that contained growth mediums: Vancomycin, EMB- lactose and PEA. They were placed in an incubator at 37º C for 1-2 days to allow growth on the mediums. The incubated microorganisms were observed to determine whether the samples were bacteria or fungi. The samples that had the bacterial colony morphology were selected for the study. Liquid cultures of the selected bacterial colony were prepared for the PCR amplification of the 16S ribosomal RNA. Two PCR tubes: tube B and tube C, were prepared with each containing 2X PCR Master Mix. Into each tube, 20 µl of 16S RNA primer mix were added. Tube B was selected to contain the prepared bacterial DNA and 5 µl of the liquid DNA was added. Tube C was the selected control tube. It didn’t contain any bacterial DNA and to make up for the absence of the DNA µl, 5µl of sterile water were added to this tube. Both tubes were brought to a total volume of 50 µl. The mixture was kept …show more content…
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
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The purpose of this lab was to identify two unknown bacteria from a mixed culture. The reason for identification of unknown bacteria was to help students recognize different bacteria through different biochemical tests and characteristics. This is important in the medical field because identification of unknown bacteria can help treat a patient by knowing the contributing source of a disease. Also knowledge of different bacteria helped others make antibiotics used today. This lab was completed by using the methods learned thus far in identification of bacteria.
The purpose of this study project was to carefully isolate and identify two unknown bacteria from a mixed culture. The ability to properly evaluate biochemical test results is also necessary for the identification to be successful. The goal was to apply all of the methods and techniques that have been learned in the microbiology laboratory course for the proper identification of unknown bacteria. A certain amount of bacteria that were used throughout the course were possible bacteria that could be found in a mixed culture. The bacteria that were identified in the mixed culture were Staphylococcus Aureus and Kocuria Rhizophila.
In a laboratory setting, it often becomes necessary to identify an unknown organism. In this experiment, researchers classified an unidentified bacterium based on its physical structure, colony morphology, optimal conditions and metabolic properties. A Gram stain using crystal violet, iodine, and safranin and a simple stain using methylene blue characterized the organism’s cell wall. Cultural behavior was classified by inoculating the organism onto nutrient agar and incubating it at 37° C for 48 hours, and observing its behavior, as well as using SIM medium to test for motility. Optimal growth temperature was
The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible
The purpose of the following study is to determine where the two unknown bacteria acquired in Microbiology lab should be classified in regards to temperature, pH level, and osmoregularity. It is important to classify bacteria in order to identify them. Identification of bacteria is important because they are not only useful but potentially dangerous as well. The identification of bacteria can lead to breakthroughs in healthcare regarding treatment of old and new diseases alike. Identifying bacteria can also be used in many other areas from better crop production through microbial pesticides to biological warfare. Their uses are endless as are their abilities to evolve and adapt to changing environments. That is why it is so important
Unknown lab report# 24 Introduction Microbiology is a study of organisms that surrounds us every day. It requires an educational understanding to identify organisms, and the ability to distinguish different bacteria’s. In applying the learning process of the different bacteria’s, unknown bacteria samples are given to be studied and identified.
The purpose of the bacterial unknown independent study experiment completed throughout the course of this lab was to determine the identity of an unknown bacterial species. The unknown bacteria sample was chosen from numerous samples provided by the instructor. The starting unknown sample, unknown #15 was a mixed bacterial culture and a broad approach taken to identify the sample. Various biochemical tests were completed to identify the bacterial species along with the use of databases such as Gideon and Bergey’s to compare the test results of known bacteria to the results of the unknown sample. Information was gathered from the other sources and databases and phenotypic testing completed and the results compared to the database results. Aseptic
Identifying microorganisms can provide information on diagnosing diseases and discovering the most beneficial treatment possible. The purpose of this assignment was to identify an unknown microorganism using biochemical tests and various methods that were practiced in the microbiology laboratory. In this paper, I will discuss the processes of how I came to identify my unknown microorganism.
The purpose to this lab was to isolate and identify two unknown bacteria from a mixed culture provided to us by our instructor. This study was done by applying all of the methods that have been instructed on thus far in microbiology laboratory class. Each test performed, provided us with some key information about the unknown microbes in question and how the bacteria function.
The objective of this experiment is to identify the organisms of two unknown bacterial cultures. Students must identify the species of the unknown bacteria by utilizing the techniques and information learned in previous laboratory exercises. These techniques include streaking for isolation, Gram staining, and specific biochemical tests. Students are given a map known as a dichotomous key, a guide in determining the identity of their unknown sample.
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.
Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)
As describes in Yamaguchi et al, the researchers purchased S. pneumoniae strain D39 from the National Collection of Type Cultures. S. pneumoniae strain R6, which is an unencapsulated derivative D39, was provided by Dr. Shin-ichi Yokota. The primers used are given. Primers are short sequences of DNA or RNA that is used as a starting point for transcription in DNA synthesis. The PCR technique is used in biology labs to create a large amount of copies of a piece of cut DNA, specifically DNA primers in this experiment(McEntee 8) S.pneumoniae and S. aureus were grown in Tryptic Soy broth or 5% sheep blood-Tryptic
After the incubation, 1.5 mL of each of the three cultures were added to eppendorf tubes and centrifuged at 13,200 rpm for 1 minute. An alkaline lysis procedure like that of Birnboim and Doly was then performed to extract the plasmid DNA with 200 μl of alkaline SDS detergent solution (Birnboim & Doly, 1979). After
Another purpose of this experiment is to stress the importance of knowing the identity of a microorganism. Knowing the species of microorganism present in a sample provides a