serve the gene sequence.
Quantifying and Observing Microorganisms 1. count/estimate the number of colonies on each plate 2. convert number of colonies using conversion factor in Table 1 to determine number of colonies per ml of solution
Antibiotic Resistance 1. Note which tetracycline plates had growth, which would indicate antibiotic resistance
Bacteria Cell Morphology Observations 1. Take a sample of bacteria from two plates of nutrient agar and two plates of agar + tetracycline 2. Make a wet mount sample of each by mixing sample with a drop of water 3. Observe wet mount sample under 10x objective and then 40x objective 4. Note cell shapes, characteristics 5. Prepare a slide for gram stain by making a wet mount but letting it air dry before use 6. Heat fix slide over a flame three times 7. Cover slide in crystal violet for 1 minute. Rinse. 8. Cover slide in Gram's iodine mordant for 1 minute. Rinse. 9. Decolorize slide in 95% alcohol for 10-20 seconds. Rinse. 10. Cover slide with safranin stain for 20-30 seconds. Rinse. 11. Dab excess water with paper towel 12. Observe slide under low objective and then 40x.
Start PCR Prep for DNA Sequence ID 1. Add colony of bacteria from plate to 100 µl water in a test tube 2. Incubate at 100 degrees C for 10 minutes and then place in centrifuge 3. Use 5 µl supernatant for PCR
-Appearance or smell of Hay Infusion Culture might vary week to week depending on the growth of bacteria in the jar.
-Colonies in plates without antibiotic have more growth, colonies in plates with antibiotic have less growth but still significant. Have large fungi growth on one of the tetracycline plates.
-Tetracycline inhibits bacterial growth in most cases; however, it is becoming more common that bacteria is resistant to tetracycline. Some of these tetracycline-sensitive bacteria include both gram-positive and gram-negative cells, such as chlamydia.
-Bacteria from sample from tetracycline plate were small circular cells, all located near each other. Bacteria from sample from nutrient agar plate were long oval-shaped cells. Looked similar to paramecium.
Conclusions The first part of this lab helped to characterize whether the bacterial growth on the tetracycline plates were antibiotic-resistant or not. Observing the cells more closely on a wet mount allowed for further analysis of the type of cell. Gram-positive and negative analysis were done on the samples from both a nutrient agar plate and from a tetracycline plate. The bacteria found on the nutrient agar plate were found to be gram negative, showing a pink color to the stain. The bacteria found on another sample of nutrient agar was found to be gram-positive, however. The bacteria found on the tetracycline plate were found to be gram-negative, and looked similar to sperillium cells. The final portion of this lab saw the beginning of DNA sequencing of the bacteria found on the plates through PCR.
Objectives 1. To understand the characteristics and diversity of plants 2. To appreciate the function and importance of fungi
The first objective will be addressed by collecting plant samples from our transects and observing/characterizing them into a major plant group. Vascularization, types of leaves, and seeds were described in order to properly characterize the type of plant. The second objective will be addressed through observing the fungi from agar plates that were preserved from the microbiology lab.
Collecting Plant Samples 1. Fill one bag with leaf litter from transect 2. Collect five different plant samples (from already dead plants)
Plant Vascularization 1. Observe the cross-section of a moss and lily slide 2. Note the vascularization of those plants 3. Note the vascularization of the plants collected from transect
Plant Specialization 1. Examine leaves of moss 2. Observe and describe the shape, size, and cluster arrangement of the leaves collected from transect
Plant Reproduction 1. Study the bryophyte reproductive cycle and identify male and female gametophytes and sporophyte 2. Dissect a lily flower sporophyte and identify the parts 3. Dissect any