Testing plant Substances as potential medicines
Purpose: To find what plant materials in our area that contain active ingredients, that will inhibit bacteria growth.
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 - pipet, 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 - pipet, 1 mL and pump - dry block heater/heat block - fine-tipped forceps - ampicillin - glass spreader - incubator oven, 37 degrees Celsius - forceps - alcohol - clamp |
Procedure:
Day 1
1. Using mortat and pestle ground up leaves with 10 ml of deionized water, let sit for 3 minutes. Filter through a filter funneland then give to teacher who will filter in a syringe and give you 1ml of your extract in a 1.7 ml tube. Label it.
2. Repeat the last step but with methanol instead of water. Label it
3. Make a negative control test with just 1m of water. Make a positive control with 1ml of ampicillin
4. Using sterile forceps drop 3 filter disks in each extract.
5. Close the tubes and let sit over night at 4 degrees celcius.
Day 2
6. Using a sterile pipet transfer 1ml of e-coli broth to the middle of the petrie dish. Sterilize a spreading loop and evenly spread the bacteria on the petrie dish.
7. Place the disks in different quadrents of the petrie dish so there are 3 of the same disks in each of the different quadrents.
8. Incubate for 24-48 hours at 37 degrees celcius.
9. Create a data table and record your results.
Day 1
1. Using mortat and pestle ground up leaves with 10 ml of deionized water, let sit for 3 minutes. Filter through a filter funneland then give to teacher who will filter in a syringe and give you 1ml of your extract in a 1.7 ml tube. Label it.
2. Repeat the last step but with methanol instead of water. Label it
3. Make a negative control test with just 1m of water. Make a positive control with 1ml of ampicillin
4. Using sterile forceps drop 3 filter disks in each extract.
5. Close the tubes and let sit over night at 4 degrees celcius.
Day 2
6. Using a sterile pipet transfer 1ml of e-coli broth to the middle of the petrie dish. Sterilize a spreading loop and evenly spread the bacteria on the petrie dish.
7. Place the disks in different quadrents of the petrie dish so there are 3 of the same disks in each of the different quadrents.
8. Incubate for 24-48 hours at 37 degrees celcius.
9. Create a data table and record your results.
Data Analysis/Conclusion
Table
Methanol Weak positive .5 ml
Water Positive 1 ml
So my plant samples showed that there are some antibodies against bacteria, as shown the water worked very well and the methanol kind of worked. Some errors could that have happened were, not using sterile equitment, human error on measureing the ammounts, and cross comtamination. We could go further by testing different bacteria on the specimins and see if the have different effects.
Table
Methanol Weak positive .5 ml
Water Positive 1 ml
So my plant samples showed that there are some antibodies against bacteria, as shown the water worked very well and the methanol kind of worked. Some errors could that have happened were, not using sterile equitment, human error on measureing the ammounts, and cross comtamination. We could go further by testing different bacteria on the specimins and see if the have different effects.
Thinking like a biotechnition
1. It does not mean that it just means that the extract doesn't work against that bacteria.
2. Yes it is a problem because the methanol could have changed its chemical compound.
3. You should try different compounds that shut down what you are trying to find.
1. It does not mean that it just means that the extract doesn't work against that bacteria.
2. Yes it is a problem because the methanol could have changed its chemical compound.
3. You should try different compounds that shut down what you are trying to find.