1. In the first leg of the experiment, one will first start by collecting the necessary materials. First, grab an empty 2% milk carton and its corresponding cap. Both the carton and its cap must be clean and dry. Along with the milk carton and cap one must get a roll of scotch tape. One the three materials are gathered that are needed for this part of the experiment one must go down to Galveston, Texas where they will fill up the milk carton with only water from one of the numerous beaches. After collecting a carton full of water, close the carton with its cap and using the scotch tape go around the cap twice. Then take the seawater carton and store it in the BSL-II lab. At the lab one must remember to feed the bacteria already existing in the sample with approximately 5 grams of fertilizer. Then reseal the carton and keep the water in an incubator with the temperature set at about 76°F.
2. For the next part of the experiment, gather another empty and clean milk carton, its corresponding cap, and a roll of scotch tape. Take the beaker and measure out 2 liters of fresh water. Next, take the empty 2% milk carton and fill it with fresh water. To the freshwater add 2 drops of the Dechlorinator. At this point, if the project is to be continued later, one must put the corresponding cap on the milk carton and seal the bottle shut with the scotch tape just as one did with the bottle of sea water. Then one must put the fresh water in the same area as the salt water, however the two containers must be placed at least a foot apart from each other.
3. The next leg of the experiment requires the Petri dish and its cap as well as the tape, the dechlorinated water, a pipette, the crude oil and the mixed liquor. From this point on the experiment will not leave the BSL-2 laboratory unless it is part of the samples that are being sent out for the TOC analyzing. Now take the container filled with mixed liquor and open it. After that take the pipette and take out 10 mL of mixed liquor. Place those 10 mL into the Petri dish. Change the tip of the pipette and then draw out 5 mL of crude oil and place that into the Petri dish as well. Last but not least fill the rest of the dish with the dechlorinated water and then place the top of the Petri dish on top and seal with tape. Keep the Petri dish in the incubator that the Sea Water should currently be residing in.
4. After a day take the Petri dish out of its incubator. Then gather the following materials: 1 of the glass jars along with its top, a pipette along with a tip, the milk carton filled with fresh water, a beaker, and the sample of crude oil. Now take the beaker and from the milk carton with fresh water measure out 1 Liter of water. Pour this water into the glass jar. Then take the Petri dish and pour its contents into the jar. Next take the pipette and from the crude oil sample measure out approximately 1 mL of crude oil and dispense it in the glass jar. Close the jar tightly and shake well for 30 seconds. Then taking a Sharpie© label the container as Fresh water. Then go ahead and place the container in the incubator.
5. Next is the control sample which requires the pipette with a clean tip, a crude oil sample, the milk carton filled with fresh water, a beaker, and a new glass container and its lid. Take the beaker and once more measure out 1 Liter of dechlorinated water/fresh water. Pour that water into the glass jar. Then take the pipette and once again measure out 1 mL of crude oil and dispense that into the glass container as well. Then close the container with its lid and shake well for 30 seconds. Finally, take the Sharpie© and label the glass container control. Then go ahead and place the container in the incubator.
6. The last sample that needs to be created is the sea water sample which requires the pipette and a clean tip along with a crude oil sample, the milk carton with sea water in it, a beaker and a new glass container with its appropriate lid. Taking the beaker measure out 1 Liter of sea water from the milk carton and pour it into the glass container. Then using the pipette measure out 1 mL of crude oil and dispense the 1 mL into the glass container. Close the container with its corresponding lid and shake well for 30 seconds. Then take the Sharpie© and label the container sea water.
7. After one day, one must revisit the samples and bring with them the 3 vials supplied by the lab along with the pipette and three disposable tips. First, one must open a sample by unscrewing the lid. Then one must take one of the pH papers and hold it in the sample for 10 minutes, after 10 minutes one must compare the colors visible on the pH to the key that analyzes what color(s) belongs to what pH level. Note the pH level of the sample. Then using the pipette they must measure and transfer 40 mL of the sample into one of the vials. Then close both the vial and the container and then put that sample back into the incubator while the vial goes inside the bubble wrap bag supplied by the lab. Once one sample is done the same thing must be done for the other two samples thus that at the end, one has three different vials each filled with a different sample as well as a pH level for each sample. All three of the vials will go inside the bubble wrap bag and the bag will go into the icebox which should always contain ice. This procedure of drawing samples and putting it into vials must be done thrice for each sample. The first time is the day after mixing the sample and then two days later another three samples must be taken and two days after that the last three samples must be taken. The pH test must be done the 2nd, 3rd, 4th, and 5th day of the experiment.
8. For the testing carbon part of the experiment one needs to take the glass vials to the Total Organic Carbon Analyzer to find the carbon content in each of the samples. A certified professional will do the testing. The machine will calculate the total amount of carbon in the sample and the experimenter must note this. After the testing is over the samples need to be disposed of as a biological agent, and the disposal must occur in the lab containing the TOC Analyzer.
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