Testing Plant Substances as Potential Medicines
Purpose: What plant materials, found locally, contain active ingredients that will inhibit the growth of bacteria?
Materials:
Balance, weigh boat, lab scoops
LB broth base
Media bottles, 250 mL
Sterilizer/autoclave
Water bath, 37*C, shaking
Sterile LB agar
Laminar flow hood and disinfectant
Plastic safety glasses
Bunsen burner and gas lighter
Inoculating loop, Ni/Cr wire
Petri dishes, 60x15mm, sterile
E. coli JM109 (stock plate)
Plant specimen
Mortar and pestle
Pipet, 10 mL and pump
Plastic funnels, short-stemmed
Filter paper disks, 5mm diameter
100 mL beakers
Syringe, 10 mL and filter, 0.2 micrometers
Reaction tubes and rack, 1.7 mL
Methanol, absolute
Pipet, 1 mL and pump
Dry block heater/heat block
Forceps, fine-tipped
Ampicillin
Glass spreader
Incubator oven, 37*C
Procedure:
Part II:
1.Weigh 2g of plant material, grind it up in the mortar and pestle along with 10 ml of de-ionized water. Let the mixture sit for three minutes, then filter through a funnel lined with paper. Then filter through a syringe to sterilize.
2.Repeat, except use 10 ml of Methanol. Put tube into heat rack to evaporate overnight.
3.Add 1 ml of filtered, sterilized water to the menthol tube and vortex it
4.Using sterile forceps, put 2 filter disks per person into each tube and leave it overnight
Part III:
1. Transfer 1 ml of E. Coli to a petri dish that is divided up into quadrants using a sterile 1 ml pipet
2. Sterilize a glass spreader with fire and brim-, I mean, alcohol.
3. Spread E. Coli throughout dish, let stand for 15 mins
4. Use sterile forceps to place discs in respective area on plate
5. Incubate at 37 Degrees Celsius for 24 hours
Materials:
Balance, weigh boat, lab scoops
LB broth base
Media bottles, 250 mL
Sterilizer/autoclave
Water bath, 37*C, shaking
Sterile LB agar
Laminar flow hood and disinfectant
Plastic safety glasses
Bunsen burner and gas lighter
Inoculating loop, Ni/Cr wire
Petri dishes, 60x15mm, sterile
E. coli JM109 (stock plate)
Plant specimen
Mortar and pestle
Pipet, 10 mL and pump
Plastic funnels, short-stemmed
Filter paper disks, 5mm diameter
100 mL beakers
Syringe, 10 mL and filter, 0.2 micrometers
Reaction tubes and rack, 1.7 mL
Methanol, absolute
Pipet, 1 mL and pump
Dry block heater/heat block
Forceps, fine-tipped
Ampicillin
Glass spreader
Incubator oven, 37*C
Procedure:
Part II:
1.Weigh 2g of plant material, grind it up in the mortar and pestle along with 10 ml of de-ionized water. Let the mixture sit for three minutes, then filter through a funnel lined with paper. Then filter through a syringe to sterilize.
2.Repeat, except use 10 ml of Methanol. Put tube into heat rack to evaporate overnight.
3.Add 1 ml of filtered, sterilized water to the menthol tube and vortex it
4.Using sterile forceps, put 2 filter disks per person into each tube and leave it overnight
Part III:
1. Transfer 1 ml of E. Coli to a petri dish that is divided up into quadrants using a sterile 1 ml pipet
2. Sterilize a glass spreader with fire and brim-, I mean, alcohol.
3. Spread E. Coli throughout dish, let stand for 15 mins
4. Use sterile forceps to place discs in respective area on plate
5. Incubate at 37 Degrees Celsius for 24 hours
Analysis & Conclusion:
My methanol results did exhibit anti-bacterial properties, however they were very weak. My water tests turned up negative. It should be noted that my negative control came out positive, so there was some experimental error. We could have had some human error, maybe we didn't break open the cells correctly, and we couldn't isolate for any one compound. Also, the ampicillin might have been diluted. If i was to do this again, I would have more data entries to make sure. However, we would also need to
Thinking Like a Bio-technician:
1. A negative result means that there are no anti-bacterial molecules in the extract.
2. That means that the alcohol killed whatever bacteria was on the disks, regardless of the extract working or not.
3. You'd have to extract and isolate each compound and test them in order to find the exact compound.
My methanol results did exhibit anti-bacterial properties, however they were very weak. My water tests turned up negative. It should be noted that my negative control came out positive, so there was some experimental error. We could have had some human error, maybe we didn't break open the cells correctly, and we couldn't isolate for any one compound. Also, the ampicillin might have been diluted. If i was to do this again, I would have more data entries to make sure. However, we would also need to
Thinking Like a Bio-technician:
1. A negative result means that there are no anti-bacterial molecules in the extract.
2. That means that the alcohol killed whatever bacteria was on the disks, regardless of the extract working or not.
3. You'd have to extract and isolate each compound and test them in order to find the exact compound.