Chemical Safety:
1. The use of all chemicals will be done while wearing gloves, aprons, and goggles.
2. All chemicals that release fumes will be placed under the fume hood to prevent the release of fumes into the classroom.
3. A fire blanket, fume-hood, fire extinguisher, eyewash station, and shower are available for use.
4. Upon leaving the lab, hands will be washed and all chemicals will be stored in a locked ventilated chemical storeroom.
Chemicals and Heat:
1. Goggles and aprons will be worn.
2. During heating, potholders (thermal gloves) will be used to handle hot surfaces and equipment coming off the hot plate.
3. No containers will be sealed when heated. In the event a closed container needs to be heated a pneumatic trough or pressure release value will be used.
1. Goggles and aprons will be worn.
2. During heating, potholders (thermal gloves) will be used to handle hot surfaces and equipment coming off the hot plate.
3. No containers will be sealed when heated. In the event a closed container needs to be heated a pneumatic trough or pressure release value will be used.
Microbiology:
Autoclave:1. Always wear goggles and aprons when using the autoclave.
2. The bottom of the auto clave should have about 2 inches of cleaned distilled water before operation.
3. Check the pressure release value prior to use (it should be clear)
4. Use potholders when opening or handling hot surfaces.
5. Opening the autoclave can be done only after the sterilizer after it has cooled (gauge should read zero.) and the all steam has been allowed to escape.
6. Place the control knob in the straight up position. This allows the unit to operate at 16-21 psi range.
Making Agar
1. Agar is to be prepared by following the proportions of dehydrated media to distilled water and then autoclaved.
2. Store sterile agar in refrigerator.
3. Excess sterile agar will be wrapped in news paper and disposed of by placing in a landfill.
Preparation of Nutrient Agar
1. Measure out 23 grams of nutrient agar dehydrated media.
2. Measure out 1.0 L of distilled water in a graduated cylinder. Place in a 2 L Erlenmeyer flask.
3. Put flask with water onto hot plate and bring to a boil.
4. Once boiling, dissolve 23 grams of agar into the water. Boil for one minute to dissolve.
5. Pour liquid agar into reagent bottles.
6. Autoclave for 30 minutes at 20psi, 120 degrees C.
7. Store in refrigerator when cooled down after autoclaving.
Culturing
1. On the underside of each petri plate label with each plate with a wax pencil.
2. Heat the nutrient agar in a microwave or water bath to 50º C.
3. Pour a thin layer of agar on the bottom of each plate and cover immediately.
4. Let agar plate solidify. Liquid agar will solidify at about 42 º C.
5. After the medium agar has solidified, streak the plate with the proper technique.
6. Once the plates have been streaked, invert and close the petri dish, tape the edges if you wish and incubate them at 25º C for 24-48 hours unless other wise noted.
Streaking Plates
1. Sterilize work area and don safety equipment
2. Take out plates from refrigerator and allow to warm up to room temperature.
3. Take out bacterial culture to be streaked onto plates.
4. Take inoculating loop and scrape off growth from bacterial culture.
5. Place inoculating loop into sterile broth of distilled water and agitate.
6. Dip sterile swab into wash.
7. Lift up lid of plate and streak wet swab onto plate being sure to cover the entire area. Plate can be streaked multiple times in a sweeping motion to ensure plate overage.
8. When done streaking plates, put into incubator upside down. Depending upon the bacteria, growth can appear as soon as 24 hours later.
9. Dispose of materials.
Cleaning up:
1. After each day of experimentation, clean the lab counter with 10% bleach.
2.Wash
3. Clean with 10% bleach all glassware and related items. Auto clave new equipment as needed.
4. Petri dishes and other disposable equipment will be placed in a plastic bag and autoclaved for 30 minutes at 20 psi, 120 degrees Celsius and disposed of.
5. All related glassware and related equipment would be autoclaved for 30 minutes at 20 psi, 120 degrees Celsius.
Disposal:1. Items of biological hazard must be disposed of after autoclaving.
2. Put on protective equipment (goggles, gloves, aprons).
3. Gather biological items to be autoclaved prior to disposal.
4. Put items into autoclavable bag. Put bag in autoclave.
5. Autoclave for thirty minutes at 20psi, 120 degrees C.
6. When able, remove from autoclave and transport to dumpster.
2. The bottom of the auto clave should have about 2 inches of cleaned distilled water before operation.
3. Check the pressure release value prior to use (it should be clear)
4. Use potholders when opening or handling hot surfaces.
5. Opening the autoclave can be done only after the sterilizer after it has cooled (gauge should read zero.) and the all steam has been allowed to escape.
6. Place the control knob in the straight up position. This allows the unit to operate at 16-21 psi range.
Making Agar
1. Agar is to be prepared by following the proportions of dehydrated media to distilled water and then autoclaved.
2. Store sterile agar in refrigerator.
3. Excess sterile agar will be wrapped in news paper and disposed of by placing in a landfill.
Preparation of Nutrient Agar
1. Measure out 23 grams of nutrient agar dehydrated media.
2. Measure out 1.0 L of distilled water in a graduated cylinder. Place in a 2 L Erlenmeyer flask.
3. Put flask with water onto hot plate and bring to a boil.
4. Once boiling, dissolve 23 grams of agar into the water. Boil for one minute to dissolve.
5. Pour liquid agar into reagent bottles.
6. Autoclave for 30 minutes at 20psi, 120 degrees C.
7. Store in refrigerator when cooled down after autoclaving.
Culturing
1. On the underside of each petri plate label with each plate with a wax pencil.
2. Heat the nutrient agar in a microwave or water bath to 50º C.
3. Pour a thin layer of agar on the bottom of each plate and cover immediately.
4. Let agar plate solidify. Liquid agar will solidify at about 42 º C.
5. After the medium agar has solidified, streak the plate with the proper technique.
6. Once the plates have been streaked, invert and close the petri dish, tape the edges if you wish and incubate them at 25º C for 24-48 hours unless other wise noted.
Streaking Plates
1. Sterilize work area and don safety equipment
2. Take out plates from refrigerator and allow to warm up to room temperature.
3. Take out bacterial culture to be streaked onto plates.
4. Take inoculating loop and scrape off growth from bacterial culture.
5. Place inoculating loop into sterile broth of distilled water and agitate.
6. Dip sterile swab into wash.
7. Lift up lid of plate and streak wet swab onto plate being sure to cover the entire area. Plate can be streaked multiple times in a sweeping motion to ensure plate overage.
8. When done streaking plates, put into incubator upside down. Depending upon the bacteria, growth can appear as soon as 24 hours later.
9. Dispose of materials.
Cleaning up:
1. After each day of experimentation, clean the lab counter with 10% bleach.
2.
3. Clean with 10% bleach all glassware and related items. Auto clave new equipment as needed.
4. Petri dishes and other disposable equipment will be placed in a plastic bag and autoclaved for 30 minutes at 20 psi, 120 degrees Celsius and disposed of.
5. All related glassware and related equipment would be autoclaved for 30 minutes at 20 psi, 120 degrees Celsius.
Disposal:1. Items of biological hazard must be disposed of after autoclaving.
2. Put on protective equipment (goggles, gloves, aprons).
3. Gather biological items to be autoclaved prior to disposal.
4. Put items into autoclavable bag. Put bag in autoclave.
5. Autoclave for thirty minutes at 20psi, 120 degrees C.
6. When able, remove from autoclave and transport to dumpster.
Electrical Power Safety
1. Limit the use of high power devices around water.
2. Never use frayed or cut wires when plugging in a device.
3. Never overload the outlet and use a surge protector.
4. All electrical devices should have an emergency cut off switch or ‘kill’ switch’.
5. Shut off and unplug all devices when not in use or when you leave the room unattended.
Power Tools
1. Wear goggles or safety glasses
2. Do not work alone in the lab.
3. Dry wet hands and clothing before working with electricity. Mop up all water spilled on the floor.
4. Keep the lab clean.
5. Clamp down any objects that will be used.
6. Know operation of all switches and controls before using a power tool.
7. Keep hands, fingers, feet, and all objects not being used way from moving parts.
8. Use a vacuum system to keep debris away from cutting area.
Sharp Object Safety
1. Use caution when using a sharp object.
2. Restrict the use of "sharps" to a minimum.
3. Do not use bent or broken needles, knifes, or razor blades.
4. Cover or cap needle after every use.
5. Discard used syringes, needles, or scalpels into an approved sharps container.
6. Bring all needles in a sharps container to a local fire station for disposal.
1. Limit the use of high power devices around water.
2. Never use frayed or cut wires when plugging in a device.
3. Never overload the outlet and use a surge protector.
4. All electrical devices should have an emergency cut off switch or ‘kill’ switch’.
5. Shut off and unplug all devices when not in use or when you leave the room unattended.
Power Tools
1. Wear goggles or safety glasses
2. Do not work alone in the lab.
3. Dry wet hands and clothing before working with electricity. Mop up all water spilled on the floor.
4. Keep the lab clean.
5. Clamp down any objects that will be used.
6. Know operation of all switches and controls before using a power tool.
7. Keep hands, fingers, feet, and all objects not being used way from moving parts.
8. Use a vacuum system to keep debris away from cutting area.
Sharp Object Safety
1. Use caution when using a sharp object.
2. Restrict the use of "sharps" to a minimum.
3. Do not use bent or broken needles, knifes, or razor blades.
4. Cover or cap needle after every use.
5. Discard used syringes, needles, or scalpels into an approved sharps container.
6. Bring all needles in a sharps container to a local fire station for disposal.
Methods:
Making the Extract;
1. Collect 5 lbs Fresh Cranberries about 3lbs
2. Put the Cranberries into the Juicer
3. Juice for 1 – 2 minutes
4. separate the juice into two equal volumes
5. Label first container (Juice Only)
6. Label second container (Fermented juice)
Fermenting:
1. Put a gram of yeast into the cranberry juice
2. Make and put into a pneumatic trough
3. Let sit for 3 weeks
Distilling the Extract;
1. Set up the Organic Chemistry Set for fractional di
2. Pour the Extract into the Round Bottom Flask
3. Turn the Heat Mantle on and set it to 60°
4. Let whatever liquid drip into a beaker for 10 minutes
5. Change the beaker
6. Raise the temperature to 70°
7. Repeat step 4 and 5
8. Raise the temperture to 80°
9. Repeat step 4 and 5
10. Raise the temperature to 90°
11. Repeat step 4 and 5
12. Collect the power at the bottom and dilute it
Inoculating the Petri Dishes;
1. Create a wash
a. Take one inoculating loop full of bacteria and swish in 5 ml of distilled water
2. Inoculating a dish
a. Dip swab into the wash
b. Make a lawn on the Petri dishes
3. Making the finished plates
a. Dip the sterile disc in the juice
b. Put the sterile disc that was dipped in the juice in a domino five formation
c. Tape up the Petri dish and label
d. Place taped Petri dishes in incubator
e. Let sit for 24 hours
f. Check the Zone of Inhibition and record
4. Collecting data
a. Check zones of Inhibition (a clear halo)
b. Measure in cm
