Synthesis Lab Report Sample

Step 1: Label the test tubes 1, 2, 3 and 4, where test tube 1 represents the sample placed on the counter at room temperature, test tube 2 represents the tube placed in the refrigerator, test tube 3 represents the tube placed in the freezer, and test tube 4 represents the sample exposed to boiling water. You will expose catalase to each of these four conditions.

This experiment was conducted to find the genus and species of an unknown bacteria prescribed by the lab teacher, which was unknown bacteria GA3 in my case. Identification of unknown bacteria techniques are used on an every day basis to figure out what type of bacteria it is and to find the best method of how to treat a patient with this bacteria (1). All five “I’s” of Microbiology were used in the testing for the unknown culture. Inoculation was used several times to put the unknown culture into agar plates or into biochemical test tubes. After Inoculation of these tubes or plates, they always were placed into the incubator for further growth and development. Isolation was used to make sure we got the correct bacteria we were testing for. After each further isolation, we gram stained the culture and inspected the culture under a microscope to further help in the identification process of the unknown bacteria. Multiple tests were done on the unknown culture to make sure we were confident in what kind of bacteria the unknown was.

Then label each of them either pGLO positive and pGLO negative, depending on which one took the plasmid (4). Place both test tubes on micro-centrifuge on ice for ten minutes. Label the agar plates with three for pGLO positive and three for pGLO negative. Label the six agar plates with what they are, whether it is just LB, LB and ampicillin, or LB, ampicillin, and araC (4). There should be one for each positive and negative. After the ten minutes in the centrifuge has passed, take them out and place them in a forty two degrees Celsius heating block for fifty seconds. Once the fifty seconds have passed, transfer the tubes back to ice for two minutes. Take the micropipette and begin adding pGLO positive or pGLO negative to the correctly labeled plates. Once the pGLO is added to the agar, use the spreader to spread out the culture until it is dry on the agar plate. Make sure that before and after every use of the spreader that it is dipped in ethanol and sterilized with the Bunsen burner, to avoid contamination. Once all of the bacteria is dried onto the agar plates, use Para film to seal the agar cultures shut. Allow for it to incubate at thirty seven degrees Celsius for twenty four hours(4). After time has passed, check for growth and use an ultraviolet light to check

Mark one tube “+” and the other “-”. The “-” tube will include the plasmid. Using a sterile graduated pipet, add 0.25 mL ice cold calcium chloride to each tube. Hold those tubes on ice. Obtain a starter plate of E. coli and using a sterile inoculating loop to transfer one large colony of bacteria form the starter plate to each tube of cold calcium chloride. In order to remove the bacteria from the transfer loop and break them apart, place the loop into the calcium chloride and twirl the loop rapidly. Dispose of the loop in a bio hazardous waste. Then, using a sterile inoculating loop, dip the loop into the DNA stock tube. When you remove the loop from the solution, check to be sure that there is a drop of liquid contained in the loop area. Transfer the liquid (approximately 10 μL to the “+” microfuge tube and twirl the loop to mix the plasmid into the solution. Place both the positive and negative tubes on ice for 15 minutes. While the tubes are on ice, obtain two Luria agar plates and two Luria agar plates with ampicillin. Label one Luria agar plate “+” and the other “-”; do the same for the Luria agar plates with ampicillin. After 15 minutes, remove the tubes from ice and immediately place them in a 42 degrees Celsius water bath for one minute. Remove the tubes from the water bath and immediately place them back on ice for two minutes. Remove them from the ice bath and add 0.25 mL of room temperature tryptic soy broth to each tube with a

3. Add 100μl of enzyme conjugate to standard wells and sample wells except for the blank well cover with an adhesive strip and incubated for 60 minutes at 37°C.

The micropipette is used to collect very small amounts of liquids that an average pipette would not be able to accurately collect. For this lab, one needs to gather two tubes which will each contain 250µL of solution. One tube will then be filled with the +pGLO plasmid and the other will contain -pGLO. The pGLO plasmid is NOT transmitted into the -pGLO tube. The solution the pGLO plasmid is in is called transformation solution and is used to soften the membrane of the E. coli cells with the calcium chloride it contains. The tubes must remain in an ice bath for ten minutes to bring their temperature down so that they will react strongly when exposed to the heat shock. Whenever researchers are transferring the E. coli bacteria from tube to tube, they must wear protective gloves and sterile loops provided to them. Researchers need to obtain and label four plates: +pGLO LB/amp, +pGLO LB/amp/ara, -pGLO LB/amp, and -pGLO LB. Amp stands for ampicillin and ara stands for arabinose. Their plates contain a base gelatin of Luria-Bertani broth. Then, once the tubes are very cold on ice, researchers place the tubes in 42-degree celsius water for fifty seconds exactly in order to activate the heat shock of the E. coli cells, prompting their membranes to become more permeable and susceptible to accepting the pGLO plasmid. After the

The first unknown test tube P found to be a gram positive bacteria. The biochemical that I performed that were positive were catalase and blood agar. Catalase has the presence of an enzyme in the test isolated detected using hydrogen peroxide. Bacteria possess catalase with a small number of bacteria; isolation is added to hydrogen peroxide when bubbles occur. When the bubbles occurred on gram positive bacterium it was a conformation of being positive. Also, blood agar plates is useful for cultivating fastidious organisms and for determining the hemolytic capabilities of an organism. There are three types of hemolysis, such as, beta- hemolysis, alpha-hemolysis, and y-hemolysis. Beta-hemolysis breaks down the red blood cells and hemoglobin completely.

Inoculated the “B12” on the blood agar plate, mannitol salt agar(MSA), phenol red mannitol/glucose broths, nutrient gelatin tube, MRVP broth, and urea agar. And then, placed everything in the “to be incubated” area. Blood agar plate helped to detect the hemolytic ability. MSA was for the salt tolerance and mannitol fermentation. Phenol red broths were for the mannitol and glucose fermentation. Nutrient gelatin tube was the test for the presence of gelatinase. Gelatin melt at 28°C; therefore, it incubated at 25°C for a week. MRVP broth incubated for 48 hours at 37°C to test for the capability of performing a mixed acid fermentation. Furthermore, it determined the ability of glucose fermentation. Urea test was for the presence of the

After the spread plate was permitted to absorb the inoculum for 10 minutes, they were inverted and incubated.

With the results of the Starch Hydrolysis Test I preceded to the final test which is the Citrate Test. Using aseptic technique, I streaked the organism onto the surface of the Simmons’ citrate slant and let it incubate overnight at 37°C. The Citrate Test determines if the organism is able to

Please refer to Dr. C. Duxbury, Department of Biology, Fall 2015, Biology 240L Fundamentals of Microbiology, Experiment 10 : Selective, Differential, and Enriched Media, pp. 53-55.

A nutrient agar plate containing the test organism with attenuated enteric bacteria was inoculated by use of a swab dipped into bacterial suspension provided. Inoculation streaks were done on the entire plate after which the agar plate was left to dry for 5 minutes. The filter paper disc impregnated with antibiotics was picked up by a pair of forceps, dipped in alcohol and passed through a flame for sterilization purposes. The filter paper disc were then pressed onto the agar plate using the forceps and incubated for 48hours at 37 degrees Celsius. The pre-incubated annual radii between the edge of the disk and the edge of the confluent were measured and the results recorded after incubation took place.

The bacteria used in this experiment is Escherichia coli which is not naturally competent. E.

Samples, cotton, sterile container, test tubes, slides, pipette, dropper, centrifuge, microscope, incubator, water bath, distilled water, 0.85% NaCl, ABH monoclonal

These streak will space out the inoculants and discrete colony of a particular specie of organism and then incubated at 35-37oC for 24hours to enhance microbial growth.