Science Fair Research Paper: Scummy Water

2. Using the tweezers, lift the steel wool out of the vinegar and shake if gently over a paper towel.

Read in your lab manual about the following agar mediums: Blood Agar (pg 168), EMB Agar (pg 170), Mannitol Salt Agar (MSA)(pg 172) ), MacConkey Agar (pg 174), and PEA Agar (pg 176) to answer the following:

room temperature. Vials 4, 5, and 6 should be in the bath containing water that is 10oC. After the

Again, label 7 1.5ml tubes 0 thru 6. Place 15μl of each serially diluted extract into its corresponding labeled tube. Next add 465μl of media into each tube. Then 60μl of Alamar blue in each tube. Finally add an additional 60μl of cells (adjusted to 10,000 cells/20 μl). Vortex each tube for 5 seconds. Now, take 3 different samples 190μl samples of concentration 0 and put it in Wells A2, B2, and C2. Repeat this step again by taking 3 more different 190μl samples of concentration 1 and putting it in wells A3, B3, C3. It should be noted that it is important to vortex each 1.5μl tube again be-fore putting it into the 96 well plate. Contin-ue this same procedure consecutively for the re-maining concentrations.

Add RO water to the 25 ml volumetric flask up to the mark. Put stopper on the flask and shake it properly.

I found that -DNA/LB was not nearly as moist as that -DNA/LB/AMP plate. The first one was dry and white.

16. Stack your group’s set of plates on top of one another and tape them together. The plates should be left upright position to allow the cell suspension to be absorbed by the agar.

After smearing the bacteria, it had to sit to air dry before being able to heat fix the bacteria onto the slide. To heat-fix, the slide was passed through the flame of the Bunsen burner 2-3 times. Once heat-fixed, the first reagent, which was crystal violet, was added to flood the slide and left on for 1 minute and then rinsed with distilled water. The second reagent, iodine, was added to the slide and left on for 1 minute and rinsed with distilled water. The third reagent, ethanol, which is also a decolorizing agent, was added to the slide for only 5-10 seconds and rinsed with distilled water.

Using the brush collect sample and put the brush in the specimen adding the brush to the sample prep and snap of the break mark and close the cap. Vortexes the sample prep for 10 seconds then remove the cap frim the heat treatment tube. Remove the dropper cap from the sample prep and squeeze 5-10 drops of the sample into the heat treatment at 95oC for 10 minutes. After remove each treatment tube and vortex for 10 seconds. Use the calibrated pipette transfer 50 microlitres from the heat mixture sample into reaction buffer tube. Replace the cap of and remove 1 test device for each sample to be tested. Open the device by pulling up and back on the large tap. Use a pipette transfer 50 microliters of the heat treated sample into the first chamber of the test device and close the cap. Check all test devices looking for any air bubbles present. Then start the amplification reaction and detection and press start. After 40 minutes result is shown if positive, it means Clostridium difficile is present.

When streaking bacteria and handling mannitol and xylitol solutions, the experimenter wore an apron, nitrile gloves on both hands and plastic protective goggles. After each set of plates were prepared, the experimenter removed the gloves from both hands without skin contact and thoroughly washed their hands for approximately 20 seconds with soap and warm water. Throughout the entire experiment the experimenter was cautious also to avoid contacting bacteria with eyes, mouth, or open wound.

When streaking with a culture swab, the swab must be first rolled and then streaked in the first quadrant of all plates. After streaking the first quadrant, an inoculation loop must be used to streak the rest of the quadrants by dragging bacteria from the previous quadrant making sure to incinerate loop between quadrants. Repeat the process for the remaining two agars.

Parts of your hands, wrists, ankles, and feet will be cleaned with an alcohol pad.

Begin by inserting a cheesecloth into a syringe and pressing the cheesecloth to the very bottom of the column.

3. Use a sterile pipette to transfer 0.1 ml of each dilution on to a MacConkey agar plate.

1. Number four clean test tubes 1-4. 2. Place 1 ml (or 20 drops) of the following solutions into each tube: water, egg albumin, starch, and chicken broth. 3. Add 3 drops of Buiret reagent to each tube and gently shake