Plant Lab:
Background:
Plants, like any other organism, have to compete with each other for resources, like water, space, and sunlight. Plants have developed different characteristics to give them advantages in competition: big leaves, height, and toxic chemicals that they excrete to kill surrounding plants. Organisms all can be infected by viruses or threatened by bacterial diseases. Some organisms have an immunity to the "foreign" invaders. Their immune system consist of antimicrobial agents that fight the microbes of the diseases. Finding and isolating the antimicrobial molecules can lead to the discovery of potential medicine. With different processes, bio-technicians extract and sample plant substances to see if they kill microbes and don't cause toxic effects in humans. After several trials, bio-technicians can determine whether the plant nutrients and chemicals can cause bacterial death.
Plants, like any other organism, have to compete with each other for resources, like water, space, and sunlight. Plants have developed different characteristics to give them advantages in competition: big leaves, height, and toxic chemicals that they excrete to kill surrounding plants. Organisms all can be infected by viruses or threatened by bacterial diseases. Some organisms have an immunity to the "foreign" invaders. Their immune system consist of antimicrobial agents that fight the microbes of the diseases. Finding and isolating the antimicrobial molecules can lead to the discovery of potential medicine. With different processes, bio-technicians extract and sample plant substances to see if they kill microbes and don't cause toxic effects in humans. After several trials, bio-technicians can determine whether the plant nutrients and chemicals can cause bacterial death.
Objective/Purspose:
To find out which plants in our area contain active ingridents that will inhibit bacteria growth.
To find out which plants in our area contain active ingridents that will inhibit bacteria growth.
Materials:
balance, weight 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 glasses, safety, plastic bunsen burner gas lighter |
inoculating loop, Ni/Cr wire
petri dishes, 60x15 mm, sterile E. coli JM109 (stock pile) plant specimen motar and pestle pipet, 10 ml and pump plastic funnels, short-stemmed filter paper disks, 5 mm diameter beakers, 100 ml |
syringe, 10 ml and filter, 0.2 um
reaction tubes and rack, 1.7 ml methanol, absolute pipet pipet, 1 ml and pump dry block heater/heat block forceps, fine-tipped ampicillin glass spreader incubator oven, 37 degrees celsius |
Procedure:
-prepare the plant extracts:
-preparing agar plates:
-prepare negative control disks (2-sterile water; 2 ampicillin):
-prepare the plant extracts:
- using a mortar and pestle, grind up two grams of plant tissue with 10mL of deionized water
- let it set for three minutes
- filter the sample through am 11 cm filter paper/funnel
- filter sterilize the extract using a syringe filter
- collect 1 mL of the filter-sterilized extract into a 1.7 microbe
- label the sample
- attach pre filters to syringe and rinse with water
- take plant extracts, syringe, prefilter, and pipet to laminar hood
- label micro fuge tube W(water) M(methonal)
- attach sterile filter to pre filter
- load 1.7 mils of extract into syringe, using pipet
- depress the plunger
- use less thank 1.0 ml filter-sterilization extract
- snap cap on micro fuge tube
- evaporte methanol from methanol extracts by placing tube with cap open on a 65 degree Celsius heat block overnight
- reconstitute methanol extract with 1.0 ml sterile deionized water
- using sterile forceps, place 3 sterile pieces of filter paper into the filtered extract
- store tubes at 4 degrees Celsius until ready to use
-preparing agar plates:
- draw a cross on each plate bottom and number the quadrants 1-4
- liquids sterile LB agar in the microwave
- using sterile technique, pour approximately 20 agar into petri plate
- using sterile forceps, add the appropriate number of sterile disks to each tube of filtered extract
-prepare negative control disks (2-sterile water; 2 ampicillin):
- place the disks into the appropriate solution
Results:
-Day 1: not enough bacteria has grown, we are going to check again tomorrow
-Day 1: not enough bacteria has grown, we are going to check again tomorrow
-Day 2: even though more bacteria has grown, our results are still inconclusive
Analysis:
We couldn't tell if the bacteria grew so our results were inconclusive. It was very hard to see the clearance.
We couldn't tell if the bacteria grew so our results were inconclusive. It was very hard to see the clearance.