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
Our results were inconclusive. Our Methanol experiment was sadly misplaced and out water experiment did not show any clearance. Just a place where the Ecoli jumped from the paper to elsewhere on the petri dish. Sadly I concluded that this "jump" was not a place where the Ecoli stopped growing, rather a place where the agar just was not there at all. Therefore the Ecoli didn't have anything to grow in or on.
Analysis
I regret to inform that this experiment could have been fun but was hardly organized. Students had to literally search through everyone else's experiment in order to even start their's. My methanol experiment and water experiment were placed in the same rack but were lost the next day. I did eventually find my water experiment after searching through 60 other experiments. However the methanol has not been found. In order for this experiment to have even happened, our main focus, and project had to be postponed until this experiment was done. In short, I believe this experiment was hardly worth the time or work needed to be put in to it.
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
- Wait and check in multiple days after completion
- This is to check for any "clearance" or area where the Ecoli does not grow
Results
Our results were inconclusive. Our Methanol experiment was sadly misplaced and out water experiment did not show any clearance. Just a place where the Ecoli jumped from the paper to elsewhere on the petri dish. Sadly I concluded that this "jump" was not a place where the Ecoli stopped growing, rather a place where the agar just was not there at all. Therefore the Ecoli didn't have anything to grow in or on.
Analysis
I regret to inform that this experiment could have been fun but was hardly organized. Students had to literally search through everyone else's experiment in order to even start their's. My methanol experiment and water experiment were placed in the same rack but were lost the next day. I did eventually find my water experiment after searching through 60 other experiments. However the methanol has not been found. In order for this experiment to have even happened, our main focus, and project had to be postponed until this experiment was done. In short, I believe this experiment was hardly worth the time or work needed to be put in to it.
Day 1 Check In: (The more Legible petri dish is the water experiment)
Day 2 Check In: (The less legible is the methanol experiment)