Metabolite extractions: Difference between revisions
(Protocols to extract metabolites from Actinobacteria) |
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==HP-20 resin Ethyl Acetate (EtOAc) extractions from liquid cultures== | |||
===HP-20 resin and Media Preparation=== | |||
HP-20 resin activation: Cover 100 g of dried HP-20 resin with approximately 300 mL of EtOAc in a clean glass bottle with polypropylene lid and shake for 10 minutes. Filter the HP-20 resin using a Buchnar funnel and filter paper and discard the EtOAc into an appropriate flammable solvent waste container. Add HP-20 resin back into the used bottle and cover again with 300 mL ethyl acetate. | |||
Adding HP-20 resin to media: Dispense 50 mL of media into 250 mL conical flasks. Use a Buchner funnel and filter paper to weigh out 5% w/v of HP-20 resin (2.5 g of HP-20 resin per 50 mL of media). | |||
Notes: the HP-20 resin should be completely dry. Never contact plastic with EtOAc or HP-20 resin that has been stored in EtOAc. Add HP-20 resin to fermentation media prior to autoclaving. | |||
Revision as of 15:28, 30 August 2019
HP-20 resin Ethyl Acetate (EtOAc) extractions from liquid cultures
HP-20 resin and Media Preparation
HP-20 resin activation: Cover 100 g of dried HP-20 resin with approximately 300 mL of EtOAc in a clean glass bottle with polypropylene lid and shake for 10 minutes. Filter the HP-20 resin using a Buchnar funnel and filter paper and discard the EtOAc into an appropriate flammable solvent waste container. Add HP-20 resin back into the used bottle and cover again with 300 mL ethyl acetate.
Adding HP-20 resin to media: Dispense 50 mL of media into 250 mL conical flasks. Use a Buchner funnel and filter paper to weigh out 5% w/v of HP-20 resin (2.5 g of HP-20 resin per 50 mL of media).
Notes: the HP-20 resin should be completely dry. Never contact plastic with EtOAc or HP-20 resin that has been stored in EtOAc. Add HP-20 resin to fermentation media prior to autoclaving.
