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BISC209/F13: Lab2: Difference between revisions
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→RICHNESS OF A MICROBIAL SOIL COMMUNITY: METABOLIC DIVERSITY: Structural Diversity in Cultured Bacteria from a Soil Community
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Directions for [[BISC209/F13: Streaking for Isolation| Streaking for Isolation]] are found in the [[BISC209/F13:Protocols| Protocols]] section of this wiki. <BR> | Directions for [[BISC209/F13: Streaking for Isolation| Streaking for Isolation]] are found in the [[BISC209/F13:Protocols| Protocols]] section of this wiki. <BR> | ||
'''Streaking for Isolation from | '''Streaking for Isolation from Solid Medium to Solid Medium:'''<BR> | ||
You can find the steps for [[BISC209/F13: Streaking for Isolation| Streaking for Isolation]] in the [[BISC209/F13:Protocols| Protocols]] section of this wiki. | |||
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<LI>Flame your loop, let it cool a few seconds. Remove or tilt the lid of the donor agar plate with your non-dominant hand but keep the lid in your hand (don't put the lid down on the bench!). With your dominant hand holding the flammed and cooled loop, touch the loop to an interesting colony of bacteria that you want to study and pick up a TINY amount of the colony. Avoid touching other bacterial colonies. Close the lid of the donor plate. | |||
<LI>Flame your loop and | <Li>Open or tilt the lid of a new sterile plate of solid medium (in this case it will be Nutrient Agar) with your non-dominant hand. Do not put the lid down on the bench; keep it in your hand! Touch the loop with the bacteria to the surface of the agar in an area near the periphery of the plate. Do not gouge the agar! | ||
<li> | |||
<LI>Flame and Repeat to streak sections 3 and 4. | <LI>Flame and Repeat to streak sections 3 and 4. | ||
<li>Invert, and incubate the plate at RT. | <li>Invert, and incubate the plate at RT. | ||
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Some bacteria often form tough leathery colonies, so transfer of these bacteria to new media to start a sub-culture is sometimes difficult. The powdery area may be spores, which would be interesting to visualize later. To isolate spore formers try to "break off" a piece of the colony with your sterile loop or with a sterile toothpick and transfer that whole piece of a colony onto zone one of the new plate. Then use your loop for streaking out the other zones. The tiny spores on the surface of the colony are likely to transfer to the next plate or tube when you work with it. (That's a good thing this time.) <BR><BR> | Some bacteria often form tough leathery colonies, so transfer of these bacteria to new media to start a sub-culture is sometimes difficult. The powdery area may be spores, which would be interesting to visualize later. To isolate spore formers try to "break off" a piece of the colony with your sterile loop or with a sterile toothpick and transfer that whole piece of a colony onto zone one of the new plate. Then use your loop for streaking out the other zones. The tiny spores on the surface of the colony are likely to transfer to the next plate or tube when you work with it. (That's a good thing this time.) <BR><BR> | ||
This process of transferring a single well isolated colony to new solid medium will continue your attempt until you succeed in isolating two different bacteria to pure culture. Our goal as a course is to find a diverse group of interesting soil bacteria . <BR><BR> | |||
The directions for [[BISC209/F13: Aseptic Transfer| Aseptic transfer: Broth to Broth and Broth to Plate]] are found in the [[BISC209/F13:Protocols| Protocols]] section of the wiki. <BR> | The directions for [[BISC209/F13: Aseptic Transfer| Aseptic transfer: Broth to Broth and Broth to Plate]] are found in the [[BISC209/F13:Protocols| Protocols]] section of the wiki. <BR> | ||
