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BISC209/F13: Lab2: Difference between revisions
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→RICHNESS OF A MICROBIAL SOIL COMMUNITY: METABOLIC DIVERSITY: Isolation & Characterization of Cultured Bacteria from a Soil Habitat
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Compare the relative number of colonies that grow on NA to the number observed on Simmons Citrate at each dilution plated from the 1% soil extract. There should be fewer colonies on the Citrate plate (A). Why? What are some important characteristics of the bacteria that are growing on this medium? <ul> | Compare the relative number of colonies that grow on NA to the number observed on Simmons Citrate at each dilution plated from the 1% soil extract. There should be fewer colonies on the Citrate plate (A). Why? What are some important characteristics of the bacteria that are growing on this medium? <ul> | ||
<LI>Count the CFU on a plate with 30-300 colonies and using a Sharpie circle and number a few of the different appearing ones on the bottom of the plate or give them code names (your initial and a number or a more creative coding scheme). | <LI>Count the CFU on a plate with 30-300 colonies and using a Sharpie circle and number a few of the different appearing ones on the bottom of the plate or give them code names (your initial and a number or a more creative coding scheme). | ||
<li>Some of the colonies on this plate may floresce. In order to see this, you will need to take a marker, safety goggles, the hand-held UV light on the instructors' desk, and your SC plate culture to a completely dark location (such as the bathroom in the hall outside the lab). Put on the glasses, turn on the UV light and shut off the white lights. If you see any colonies that "light up"(B), | <li>Some of the colonies on this plate may floresce. In order to see this, you will need to take a marker, safety goggles, the hand-held UV light on the instructors' desk, and your SC plate culture to a completely dark location (such as the bathroom in the hall outside the lab). Put on the glasses, turn on the UV light and shut off the white lights. If you see any colonies that "light up"(B), mark them with a small dot or circle on the bottom side of the plastic plate and attempt to isolate at least one of them to pure culture. <BR> | ||
<li> Make observations in your lab notebook about the #, appearance, color, consistency or florescence of any colonies growing on your Simmons Citrate plates. | <li> Make observations in your lab notebook about the #, appearance, color, consistency or florescence of any colonies growing on your Simmons Citrate plates. | ||
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<li>Follow the directions found in [[BISC209/F13: Streaking for Isolation| Streaking for Isolation]] in the [[BISC209/F13:Protocols| Protocols]] section of this wiki carefully. Transfer bacteria from this colony to the O zone of a new NA plate. | <li>Follow the directions found in [[BISC209/F13: Streaking for Isolation| Streaking for Isolation]] in the [[BISC209/F13:Protocols| Protocols]] section of this wiki carefully. Transfer bacteria from this colony to the O zone of a new NA plate. | ||
<li> Flame and cool your loop before going back into sequential, inoculated zones to transfer fewer and fewer bacteria as you streak each zone. | <li> Flame and cool your loop before going back into sequential, inoculated zones to transfer fewer and fewer bacteria as you streak each zone. | ||
<LI>Parafilm and save any plate that contains a colony you or your teammates are attempting to isolate. Discard | <LI>Parafilm and save any plate that contains a colony you or your teammates are attempting to isolate. Discard plates that you do not contain colonies that were subcultured. | ||
<li> Check the sub-cultures frequently and sub-culture over and over to new media until you are sure that your culture is pure. Note how fast your organism grows and if the initial colony appearance and/or fluorescence is maintained. | <li> Check the sub-cultures frequently and sub-culture over and over to new media until you are sure that your culture is pure. Note how fast your organism grows and if the initial colony appearance and/or fluorescence is maintained. | ||
