Materials
- Balance, weight boat, lab scoops
- LB broth base
- Media bottles, 250 mL
- Sterilizer/Autoclave
- Water bath, 37 degrees Celsius
- Sterile LB agar
- Laminar flow hood and disinfectant
- Plastic safety glasses
- Bunsen burner and gas lighter
- Inoculating loop, Ni/Cr wire
- Petri dishes, 60 x 15 mm, sterile
- E. coli JM109 (stock plate)
- Plant specimen
- Mortar and pestle
- Pipette, 10 mL and pump
- Short-stemmed plastic funnels
- Filter paper disks, 5 mm diameter
- Beakers, 100 mL
- Syringe, 10 mL and filter, 0.2 microliters
- Reaction tubes and rack, 1.7 mL
- Absolute methanol
- Pipette, 1 mL and pump
- Dry block heater/heat block
- Fine-tipped forceps
- Ampicillin
- Glass spreader
- Incubator oven, 37 degrees Celsius
- Forceps
- Alcohol
- LB broth base
- Media bottles, 250 mL
- Sterilizer/Autoclave
- Water bath, 37 degrees Celsius
- Sterile LB agar
- Laminar flow hood and disinfectant
- Plastic safety glasses
- Bunsen burner and gas lighter
- Inoculating loop, Ni/Cr wire
- Petri dishes, 60 x 15 mm, sterile
- E. coli JM109 (stock plate)
- Plant specimen
- Mortar and pestle
- Pipette, 10 mL and pump
- Short-stemmed plastic funnels
- Filter paper disks, 5 mm diameter
- Beakers, 100 mL
- Syringe, 10 mL and filter, 0.2 microliters
- Reaction tubes and rack, 1.7 mL
- Absolute methanol
- Pipette, 1 mL and pump
- Dry block heater/heat block
- Fine-tipped forceps
- Ampicillin
- Glass spreader
- Incubator oven, 37 degrees Celsius
- Forceps
- Alcohol
Procedure
- Grind up 2 g of plant leaf material using a mortar and pestle with 10 ml of deionized water.
- Filter the sample through a 11 cm filter paper funnel.
- Filter sterilize the sample extract using a syringe filter.
- Collect 1ml of extract into a 1.7 ml microtube.
- Label the sample.
- Repeat this last step, but using methanol instead of water.
- Place 1 ml of methanol extraction into a 1.7ml tube and place the tube with the cap open into a heat block set at 65 degrees F.
- Using sterile forceps, drop three filter paper discs into each tube filled with extracts.
- Let sit overnight at room temperature until ready to test.
Results
After conducting this experiment, we found that the areas around our plant samples were lacking bacterial growth. This shows that some substance in our plant samples inhibits bacterial growth, or kills bacteria.