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Autoclave Machine Lab Report

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actococcus lactis ssp. lactis C2 and bacteriophages c2, ml3, sk1 were obtained from T. R. Klaenhammer (North Carolina State University, Raleigh, NC). Both bacterial Lactococcus lactis ssp. lactis C2 stains and bacteriophages c2, ml3, sk1 were stored in microcentrifuge tubes at -80℃ refrigerator to keep frozen until use.

3.3 Preparation of M17 medium, skim milk, Bottom agar, Top agar and CaCl2 solution M17 medium, skim milk, Bottom agar, Top agar and CaCl2 solution were all prepared in lab 201 in Garrugus building (University of Kentucky). All the materials and solutions involved in this research were autoclaved for 15 minutes at 121℃ in microbiology lab 202 (University of Kentucky). The complete cycle in the autoclave machine was about …show more content…

Top agar was dispensed into 50ml test tubes and then autoclaved for 15 minutes at 121℃ as well. Stored the top agar at -5℃ refrigerator when it cooled down. Preheated top agar for 20 minutes to liquefy before use. The optimal dispense temperature for top agar was 45℃.

3.3.4 Preparation of CaCl2 solution The molecular weight of CaCl2 was 111 g/mole. Weighed 11.1g CaCl2 and dissolved into 100ml distilled water. The final CaCl2 concentration would be 1M. CaCl2 solution was used to make bottom agar and initiate the infection cycle.

3.4 Selection of most sensitive strain to bacteriophages to make new stock culture Starting culture was prepared by inoculating 1ml (1×109 CFU/ml) stock Lactococcus lactis ssp. lactis C2 culture into 25ml M17 medium in a test tube. The mixture was incubated overnight for 16h in a 32℃ water bath to get fresh Lactococcus lactis ssp. lactis C2 culture. Inoculated the Lactococcus lactis ssp. lactis C2 culture onto the petri dish with bottom agar by using inoculation loop for streaking. The streaking was used to isolate the pure strain of Lactococcus lactis ssp. lactis C2 from the stock culture. The inoculation loop was sterilized in a fire for 5-7 seconds and then cooled to dip into the starting culture for microbes inoculation on the plate. Allowed the inoculation plate to grow in a constant 32℃ temperature incubator for another 16 hours. After 16 hours, the plate we streaked had shown colonies thinning with the streaking direction

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The purpose of this lab report is to employ a myriad of skills, tools and, methods learned throughout this semester to perform the appropriate tests for the identification of the assigned unknown bacteria. Add more background information here!!! The most important tools and techniques used during this identification include aseptic technique, microscopic examination and, the use of selective and differential media. Aseptic technique is an important tool for microbiologists. It is imperative that aseptic technique is maintained throughout the length of any test to avoid any cross-contamination that may lead to inaccurate results.

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The unknown bacteria was then tested on multiple selective and differential media. Growth was present on the MacConkey Agar and the colonies were the same color as the plate, which told me my bacteria was gram negative and did not ferment lactose. There was no growth on the Mannitol Salt Agar, and this told me the unknown was not salt tolerant and did not

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Identification of bacteria within Unknown Culture #21 In this experiment, an unknown culture of two different types of bacteria was assigned to each person, a number of tests were performed to isolate and identify these bacterial cells. Based on knowledge from the previous experiments completed in lab, a basic understanding of each type of bacteria was used to create a flow chart that would aid the process of identifying the unknown bacteria within the culture. A gram stain that is performed initially will narrow down the types of tests certain bacteria will and will not respond to. In addition to the gram stain, some of the tests that were used include, a catalase test, an Eosin methylene blue (EMB) agar test, a bile esculin test, and a 6.5% sodium chloride (NaCl) test.

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The tube was placed back in incubation for 96 more hours to observe any more positives. 2.10 Catalase Test A trypticase soy agar plate was used and after incubation, four drops of 3% Hydrogen Peroxide was added to the plate to flow over the bacterial growth. A presence of bubbling was observed. 2.11 Starch Hydrolysis

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Transformation was successful in the plates where the bacteria consumed the pGLO plasmid. In the first plate that the bacterium was plated on it included the LB broth and of ampicillin antibiotic (amp), 2 colonies were present. The second plate of bacteria was grown with the presence of LB broth, ampicillin, arabinose sugar (ara), and 22 colonies were observed. But a green fluorescent glow of the colonies was only present in plate 2. Plates 3 and 4 were the control plates.

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Aseptic technique was initiated at the beginning of this experiment by cleaning the work surface with disinfected wipes. Personal protectives equipment was also worn. The material utilized in this experiment was: S. epidermidis culture broth, sterile cotton swab, streak plate, forceps in 70% alcohol, a lit tea light, and the three antibiotic disks (novobiocin, gentamicin, penicillin). The first step, I divided a plate into three quadrants and labelled them with the different antibiotic names. Using the lit tea light, like a bursen burner, I flamed the mouth of the S. epidermidis culture.

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The B. Vulgaris samples were approximately 1cm3. They were kept the same size to ensure accurate results. A control test was conducted in distilled water to obtain a result to compare. The ethanol treatments were 40% and 70%. To prepare the solutions a 70% ethanol solution was used to make 40%.

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The culture was maintained on a nutrient agar plate/slants at 4°C and as glycerol stocks 40% at −70°C. Preparation of bacterial supernatant for synthesis of nanoparticles A loopful of the culture was inoculated into 250 ml sterile nutrient broth and incubated at 37°C for 24 hours. The

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The Another medium used was MAC, it is used to isolate and differentiate gram-negative organisms and it is a pink, dusty rose color. Lastly, the Citrate Slant is a green color and it was used as a differential test to examine enzymes. The media were inoculated at 37°C for 48 hours, then it was observed to determine the

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There are multiple points both at 43°C and at 72°C which indicates that liquid was collected at these temperatures. Based on this information, it would appear that two different liquids were present in solution and that one liquid has a boiling point of approximately 43°C and that the other has a boiling point of approximately 72°C. The literature value boiling point for DCM in is reported to be about 40°C and it is about 80°C for cyclohexane. Based on the graph, DCM was collected from 4 ml to 22 ml, thus 18 ml of DCM was collected.

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The organism that was successfully cultured from the first lab was use to streak into other plates. Samples from the previously cultured MAC and blood agar were streaked on two different sides of an EMB, SS, and MSA plate. Samples of bacteria were also used to inoculate Citrate and TSI media. The inoculation of TSI and Citrate media were as follows: The materials were gathered, which include the previously cultured media, an inoculation needle and the Citrate and TSI medium.

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The developing solution was poured into a tank and was tightly covered with a glass lid, and the tank was allowed to be saturated to ensure that the solution was equilibrated in the gas phase. Silica plate for TLC analysis: A horizontal line was drawn with a pencil on the plate and it was about 1 cm above the bottom of the plate. The horizontal line was drawn faintly so as to avoid damaging the silica gel on the plate. On the horizontal line, two marks were made and one was named A and the other B. These marks were made towards the centre of the plate at a distance apart because when spots are made at the edge of a plate, the result would be an improper travel of the samples as the solvent advances on the plate.

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