BISC314:Full Protocol: Difference between revisions
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'''LAB #3: Antagonistic and Mutualistic Interactions'''<br> | '''LAB #3: Antagonistic and Mutualistic Interactions'''<br> | ||
**NOTE** You must come in the day prior to set up overnight cultures of your isolates!!!<br> | **NOTE** You must come in the day prior to set up overnight cultures for 8 of your isolates!!!<br> | ||
The microbial community living within the cheese is a complex one with many different microorganisms. As is true of any environment, these microbes interact with each other - both metabolically and chemically. Today, you will be taking your isolates and testing them against each other for growth inhibition or growth benefit. Some of these organisms may prevent the growth of others through the production of chemicals we'll discuss in lecture; others might promote the growth of their neighbors by secreting metabolites that are needed. <br> | The microbial community living within the cheese is a complex one with many different microorganisms. As is true of any environment, these microbes interact with each other - both metabolically and chemically. Today, you will be taking your isolates and testing them against each other for growth inhibition or growth benefit. Some of these organisms may prevent the growth of others through the production of chemicals we'll discuss in lecture; others might promote the growth of their neighbors by secreting metabolites that are needed. <br> | ||
<b> Interaction Assay </b> <br> | <b> Interaction Assay </b> <br> | ||
1. Create a 96-well template for your isolates. Using the excel template found [[Media:template.xls]]. Using sterile technique, take | 1. Create a 96-well template for your isolates. Using the excel template found here [[Media:template.xls]] make up the map for this 96-well plate - you should know where each of your 8 chosen isolates will be. <br> | ||
2. Using sterile technique, take 50 ul of your isolate from the overnight culture and place it into the appropriate well. Add another 100 ul of sterile media to the well to dilute the culture <br> | |||
3. When you are finished loading the top and side wells, use the p 20 micropipette to move 10 ul from wells A2 - A8 into wells B2 - B8. Repeat this until H2 - H8 has 10 ul in it also. <br> | |||
4. Now we will add the organisms from wells B1 - H1 to the rest of the plate. Again, using the p 20, take 10 ul from wells B1 - H1 and mix into the wells B2 - H2. Repeat this process until you have mixed all of the possible combinations of your isolates on this plate. <br> | |||
5. Each of your wells now has isolates growing by themselves and isolates mixed together. We will inoculate from this plate onto a square agar tray in order to observe what these organisms look like when grown together. For this we will use the multichannel micropipette. | |||