Testing plant substances for medical purposes lab/reflection
Anthony Allen & Jack Tarleton
Background-
Plants are in a constant race for light and food. Certain characteristics such as fast growth, big leaves, and large roots prove to be very advantageous. Some plants have developed toxic chemicals that fall from their leaves, into the soil, and kill the competing plants. Many plants develop more of a defense such as antimicrobials to battle foreign viruses and bacteria. Identifying the microbes you are looking for is not trivial. Out of the millions of plants, each with millions of different compounds many samples take a very long time to find. A certain plant may even be somewhere far away, such as the jungles of brazil.
Purpose-
What plant materials, found locally, contain active ingredients that will inhibit the growth of bacteria?
Materials-
Balance, weight boat lab scoop, LB broth base, media bottles, sterilizer,/autoclave, water bath, sterile LB agar, laminar flow hood and disinfectant, safety glasses, Bunsen burner and gas lighter, inoculating loop, Ni/Cr wire, petri dishes((60x15mm)), E.Coli JM109, plant specimen, mortar and pestle, pipet((10 mL and pump)), plastic funnels, filter paper discs, 5mm diameter beakers, syringe, reaction tubes, methanol, absolute, pipet((1 mL and pump)), dry block heater/heat block, forceps ((fine tipped)), ampicillin, glass spreader, incubation oven ((37 degrees celsius))
Procedure-
Results-
The results of this project were not what we completely thought they would be. We expected to see a large amount of ecoli sprouting around our paper, yet we did not get these results. We had 2 experiments, methanol and water, the methanol experiment was misplaced, yet the water experiment did show a little bit of results. The other groups that we collaborated with had a bunch of ecoli show up on their experiment, but we had only a small amount that came about just around the paper experiment. Going in depth, the project goal was not reached in our group. If we did this again I would have kept better watch on my experiment instead of lose it, which nobody is to blame but myself. Official scientists do the same thing at times, my mistake did not ruin the whole experiment, it just diverged my results and that is just a lesson to learn, live and learn.
Background-
Plants are in a constant race for light and food. Certain characteristics such as fast growth, big leaves, and large roots prove to be very advantageous. Some plants have developed toxic chemicals that fall from their leaves, into the soil, and kill the competing plants. Many plants develop more of a defense such as antimicrobials to battle foreign viruses and bacteria. Identifying the microbes you are looking for is not trivial. Out of the millions of plants, each with millions of different compounds many samples take a very long time to find. A certain plant may even be somewhere far away, such as the jungles of brazil.
Purpose-
What plant materials, found locally, contain active ingredients that will inhibit the growth of bacteria?
Materials-
Balance, weight boat lab scoop, LB broth base, media bottles, sterilizer,/autoclave, water bath, sterile LB agar, laminar flow hood and disinfectant, safety glasses, Bunsen burner and gas lighter, inoculating loop, Ni/Cr wire, petri dishes((60x15mm)), E.Coli JM109, plant specimen, mortar and pestle, pipet((10 mL and pump)), plastic funnels, filter paper discs, 5mm diameter beakers, syringe, reaction tubes, methanol, absolute, pipet((1 mL and pump)), dry block heater/heat block, forceps ((fine tipped)), ampicillin, glass spreader, incubation oven ((37 degrees celsius))
Procedure-
- Preparing plant extracts:
- Use a mortar and paste to grind up 2 grams of plant tissue with 10 ml of deionized water
- Let it sit for 3 minutes
- Filter the sample through an 11 cm filter paper/funnel
- Filter/sterilize the extract using a syringe filter
- Collect 1 mL of the filter-standard extract into a 17 microtube. Label the sample
- Attach prefilter to syringe and rinse with water
- Take to Laminar hood: Plant extract, Syringe/prefilter, Microfuge tube rack, Pipet
- Label microfuge tube with initials and whether it is water/methanol and put in rack
- Attach the sterile filter to the prefilter
- Load 1.7 mL of extract to syringe using pipet
- Put plunger in and depress it
- Have at least 1 mL of sterilized extract
- Snap on cap on microfuge tube
- Evaporate methanol from methanol extracts by placing a tube, with a cap, upon a 65 degree heat-block overnight
- Reconstitute methanol extract 1 mL sterile deionized water
- Using sterile forceps place 3 sterile pieces of filter paper into the filtered extract 4 degree celsius
- Store until ready to use
- Draw a + on each plate bottom and number quadrants 1-4
- Liquify sterile LB agar in the microwave
- Using sterile technique pour approximately 20 mL of agar into Petri plate
- Using sterile forceps add the appropriate number of sterile disks to each tube of filtered extract
- Label both plates with either M for methanol or W for water
- Place the disks into the appropriate solution
Results-
The results of this project were not what we completely thought they would be. We expected to see a large amount of ecoli sprouting around our paper, yet we did not get these results. We had 2 experiments, methanol and water, the methanol experiment was misplaced, yet the water experiment did show a little bit of results. The other groups that we collaborated with had a bunch of ecoli show up on their experiment, but we had only a small amount that came about just around the paper experiment. Going in depth, the project goal was not reached in our group. If we did this again I would have kept better watch on my experiment instead of lose it, which nobody is to blame but myself. Official scientists do the same thing at times, my mistake did not ruin the whole experiment, it just diverged my results and that is just a lesson to learn, live and learn.
Analysis-
The project that we worked on was definitely a very different project than I've ever done. We grabbed a few ounces of plant and tried to find if they had anything that could be used as a bacterial fighting medicine in a way. The only thing that I disliked was how we lost our project, my group mate and I looked everywhere for our methanol experiment, but to no avail. The best thing was that we took something from some of our backyards and turned it into an experiment that could possibly help people
in need. I would want to do this again to try and find out the difference from the water and methanol experiments and find out the exact result instead of having in conclusive results. I would rate this project a 7/10 due to the fact that I just misplaced my experiment. That is what I thought about the project.
The project that we worked on was definitely a very different project than I've ever done. We grabbed a few ounces of plant and tried to find if they had anything that could be used as a bacterial fighting medicine in a way. The only thing that I disliked was how we lost our project, my group mate and I looked everywhere for our methanol experiment, but to no avail. The best thing was that we took something from some of our backyards and turned it into an experiment that could possibly help people
in need. I would want to do this again to try and find out the difference from the water and methanol experiments and find out the exact result instead of having in conclusive results. I would rate this project a 7/10 due to the fact that I just misplaced my experiment. That is what I thought about the project.