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Phage Lab Report

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Collecting an Environmental Sample: A soil sample was obtained from the greenhouse in Abelson Hall on the seventh floor at Washington State University. The coordinates of the site are 46.729584, -117.165361. The sample was wet, and the temperature was 24 degrees Celsius. Isolating a novel phage from the environment: The purpose of this experiment was to extract phage from the sample to infect the host bacteria. This protocol shows the variety of phages in the sample. 3-5ml of the sample was poured into the bioreactor enrichment medium. Next, .5ml of M. foliorum was poured into the bioreactor, closed, labeled, and put in a shaker for 48 hours. Enriched Isolations: Enriched isolation was used to amplify phages present in the sample. This protocol allows bacteria to grow, replicate, infect, and form a large mass of phage. 1ml of the enrichment and soil mixture was transferred to a microfuge …show more content…

This procedure requires plaque isolation, which enables characterization of a single phage population. A single plaque was chosen from the plate by circling the plaque with a sharpie and touching it with a sterile wooden applicator. The plaque was aseptically streaked onto a new agar plate. The wooden applicator was disposed, and a new applicator was obtained to start a new streak, starting from the end of the previous streak and repeated for a total of 3 streaks on the plate. The end of the third streak was marked with an “X” using a sharpie. The plate was then properly labeled. To prepare the agar plates, 20ul of CaCl2 was added to a tube of top agar. Next, a tube of top agar was poured into a tube of M. foliorum and mixed gently. The mixture in the last step was then poured onto the L-agar plate where the “X” mark was made. After sitting for 10 minutes, the plate was incubated in the 30-degree Celsius incubator for 48 hours. The experiment will continue next lab by performing pure phage

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Then they are shocked with a calcium chloride solution that changes the charge on the cell membrane so that the plasmid DNA may be accepted into the cell. This solution must be chilled so that the cell membrane may heal. After incubating the bacterial cells, they are heat shocked to open the pores in the cell membrane to allow the transformation to occur. After being chilled again in order not to melt the agar, the cells are placed in a medium

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When bacteria are allowed to replicate in a petri dish and the temperature is at an optimal 37 degrees Celsius, normally cloudy patters form on the agar similar to frost on a windowpane. D'Herelle instead observed small, clear, perfectly circular spots that had formed amid the bacterial growth. He tucked away this newly acquired knowledge, and years later he discovered his proof that the phenomenon of the clear spots wasn't limited to the coccobacilli of locusts, but that bacteria pathogenic to man were equally susceptible to it when testing human stool from soldiers potentially with dysentery. To analyze what were killing the bacteria,

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The agar did not change color from pink to yellow, meaning that no mannitol fermentation occurred. After comparing the result of no mannitol fermentation to Bergey’s Manual, as well as the other test results, it was possible to conclude that the gram positive bacteria was Micrococcus

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Introduction: Microbiology is the study of the microbial world, which is composed of microorganisms that are too tiny to be seen with a naked eye. This project demonstrates the significance of correctly identifying unknown organisms with methods and techniques acquired in the laboratory as they can make or break one's ability to accurately isolate and identify various species. Studying microbes enables oneself to have an insight on not just their structure or how they function, but how they have an impact on organisms and their environment around them as they can be found nearly anywhere. The importance of the process of identifying unknown bacteria vary from determining the causative agent of a disease, to testing if a specific food or fluid is safe to consume, and simply knowing what is present

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Beginning in September of 2020, as a phage researcher in the SEA-PHAGES program under the supervision of Dr. Susan Gurney and Dr. Brett Condon, I worked with other members of the team to isolate a bacteriophage that had never been discovered. I then utilized bioinformatics to study the virus’ DNA sequence, and this culminated in a poster presentation at the National SEA-PHAGES symposium as well as a publication titled “Isolation and characterization of Microbacterium phages through a virtual lab experience. " This experience taught me the importance of meticulous attention to detail in research, and I had the honor of learning how scientific experiments are performed by two distinguished and knowledgeable professors. I will work hard to apply this same attention to detail to my time as a

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Gel is checked in 20 minutes to determine how far blue dye has travelled. Gel is carefully removed (with gloves) and placed under UV light to take pictures. Phage Cluster Identification and Agarose Gel Electrophoresis ll 20µL of sterile water and 5µL of phage genomic DNA is added to tube. Diluted DNA (2µL) was added to each PCR tube. Tubes was placed in thermo cycle and program started.

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We then flipped the dish and sectioned it off into 4 sections, which then were marked with the specific genotype that would be inoculated into that section. The initials of the group were also put on the dish. Then we used an inoculating loop to cut out sections of the fungus. The inoculating loop was sterilized with a flame and let cool down before touching the fungus. After cutting a block of the fungus, we placed it on the petri dished in the section that was appropriately marked for that specific strain.

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Erica L. Maul. Rachel Richman Microbiology 485 27 March 2024 Prospectus With the objective of demonstrating the knowledge and skills I have acquired in the Microbiology Program at NDSU, I will be aiding in research in the Geddes Lab for my Capstone Experience. Under the guidance of Dr. Barney Geddes, alongside the mentorship of graduate student Natalie Visich, I anticipate a valuable experience full of learning. In particular, I am curious about the pace of research.

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Materials Based off of the flow chart and the methods took to identify the two unknowns the tests I conducted were the Gram Stain Test, Endospore stain test, Urease test, Lactase test, and lastly the MRVP Methyl Red test. The first test conducted was the Gram Stain test. This test was used to determine if the two bacteria were either Gram Negative or Gram Positive. There are four main steps in the gram stain process that all affect the outcome and overall stain and accuracy of the test. The Primary stain is the first step in the gram stain.

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The purpose of this experiment is to measure the concentration of virus in

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The plates were then stored at 32°C (90°F) for one week to allow the bacteria enough time to have growth. My lab Partner Kat Lohmeyer and I measured the different colonies formed in each sample, along with the different morphology types. Introduction Prokaryotic

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Gram-staining was done by drying the bacteria then staining it with Crystal violet. After that, iodine was applied, then a layer of Safranin was applied. If the crystal violet is still visible, the bacterium is Gram-positive, but if the bacterium is closer to pink, it is Gram-negative. The second step of the experiment was growing the bacteria. In this section, Bacteria F was streaked across the agar plate.

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Arguments against the use of Phages are numerous, but all of them are based around the beginning of phage use. At this time, solutions to these problems have already been found. Issue one is based on the host range of phages. A microbiologist named Michael Schmidt at Medical University of South Carolina described the range of antibiotics and phages as: “Antibiotics are a big hammer... You want a guided missile.”

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I was awarded a position in my university’s research program under mentorship of Dr. Kirk Anders in yet another enriching research experience studying one of my most prominent passions- bacteriophage. Matching Dr. Ander’s visions, this experience felt like an authentic independent research project. I began in the spring of my junior year and continued through the summer months and into my senior year. After burying myself in the lab’s literature and soaking up my mentor’s knowledge of promoter activity in bacteriophage, I was able to complete two projects concerning the activity of predicted promoters in a bacteriophage. Initially, I struggled to make any significant progress, as shown through my many unsuccessful PCR reactions and baffling sequencing results; however, by the end of my first semester of research, everything began to work and the results came flooding in.

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In this lab, the quantitative assay that was constructed was a plaque assay. A plaque assay is a modification of a bacteriophage assay and is used to estimate the titer (concentration of a solution) of a phage stock. A plaque assay contains a virus combined with bacterial cells on the surface of an agar plate. The agar plate inhibits the spread of viral progeny to neighboring bacterial cells which results in plaque formation (lysis of bacterial cell). The purpose of this lab was to perform a bacteriophage assay and to calculate the original stock titer of the phage stock using the known set of dilutions and direct plaque count.

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