Unknown Mixed Microorganisms

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 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

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 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

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 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.

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

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.

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)

This laboratory experiment’s objective was to take a pure culture and isolate it from a mixed culture. The other part of the objective was to ascertain what species of bacteria that the pure culture was. The hypothesis made stated that so long as lab protocol was followed, the unidentified culture would be positively recognized/identified. An isolated pure colony of the unknown culture was obtained using the quadrant streak plate method. Afterward, the culture was Gram stained, and the results showed that it was Gram positive. Motility tests were done on the unknown using a filter paper bridge on a petri dish that contained TTC with agar. The unknown was revealed to not be motile, which meant that it did not possess flagella. The last test done was to learn the metabolic capabilities of the unknown bacteria. There were tests done for citrate utilization, the mixed fermentation pathway, catalase presence, carbohydrate fermentation in mannitol, lactose and glucose, urease production and the butanediol fermentation pathway in order to better identify the unknown bacteria. The results from each of the metabolic tests in conjunction with the motility and Gram staining tests were ultimately compared to results from database containing many different kinds of results from various bacteria. The unknown from the mixed culture was identified as Staphylococcus

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

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.

When the mixtures ready and added to Applied Biosystems wells we have run them to PCR machine as these conditions activation at 50 °C for 2 minutes and at 95°C for 2 minutes and Amplification at 95 °C for 1 seconds; 60 °C for 30 seconds × 40 (Applied Biosystem, 2010).

Sediment DNA was extracted using the Powersoil DNA extraction kit ((MoBio Laboratories Inc., USA) following manufacturer's protocol. Recovered DNA was quantified using QuBit meter using QuBit dsDNA HS kit (Life Technologies, Portugal). Different primer pair specific to hyydrazine odi…… gene (HZO gene) was used to amplify DNA of anaerobic ammonia oxidizing bacteir (anammox). Due to low annealing temperature, the amplified product was non-specific. Later with primer (ref…) with slight modification HZO gene of anammox bacteria was amplified in a PCR reaction containing 1X EmeraldAmp® GT PCR Master Mix (Takara Bio Inc. Japan), 1mM primer each and 20-30 ng of soil DNA using the programme as follows: initial denaturation of 95°C for 5 min followed by 42 cycles of denaturation at 95°C for 60 s, 60 °C of annealing for 60 s, extension of 75°C for 2 m and a final extension of 75°C for 5 min. The pooled amplified product was ligated to pGEM-T easy vector (Promega, USA) and transformed into DH5α E. coli competent cells. The clone library was prepared from the positive transformants, and the clones were preserved using 60% glycerol at -80°C. Ninety-six positive clones were selected for plasmid isolation using alkaline lysis with resuspension

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