Difference between revisions of "Antimicrobial Peptide Extraction"
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-Discard the supernatant and let the protein precipitate dry overnight | -Discard the supernatant and let the protein precipitate dry overnight | ||
| − | -Wash the protein out of the centrifuge tubes with 35% Acetonitrile into a plastic tube | + | -Wash the protein out of the centrifuge tubes with 35% Acetonitrile and a plastic pipet into a plastic tube |
===Mutacin 1140 extraction=== | ===Mutacin 1140 extraction=== | ||
Revision as of 12:17, 14 July 2015
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Contents
Purpose
This protocol is used for extracting antimicrobial peptides made by bacterial species.
Materials Needed
Occidiofungin extraction
-Potato dextrose broth
-Burkholderia contaminans
-large centrifuge tubes
-centrifuge
-35% Acetonitrile
-2000 mL flasks
-stir bar
-stir plate
-Sodium Thiosulfate
-plastic pipette
Mutacin 1140 extraction
Protocol
Occidiofungin extraction
-Culture the Burkholderia contaminans bacteria in Potato Dextrose Broth at 25 C for at least 5 days
-Centrifuge the bacteria at 13,000 rpm for 20 minutes
-Pour the supernatant into 2000 mL glass flask with a stir bar and add 0.5 volume of Sodium Thiosulfate (i.e. 750 mL of supernatant, 375 grams Sodium Thiosulfate)
-Stir the mixture for 2 hours
-Centrifuge the mixture at 13,000 rpm for 20 minutes
-Discard the supernatant and let the protein precipitate dry overnight
-Wash the protein out of the centrifuge tubes with 35% Acetonitrile and a plastic pipet into a plastic tube
Mutacin 1140 extraction
Notes
Papers where this or a similar method has been used
Related Ontology Terms
References
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