Actinomycete library preparation and large scale cultivation
Faculty Mentor Name
Dr. Skylar Carlson
Research or Creativity Area
Natural Sciences
Abstract
Actinomycetes are a phylum of gram-positive bacteria known to dedicate a large portion of their genome to secondary metabolite production. These secondary metabolites are small, drug-like molecules that have potent biological activity useful in inspiring new drug candidates. The Carlson Lab Actinomycete Collection is composed of approximately 100 strains. These have been isolated from the Great Lakes freshwater sponges, Puerto Rico sediments, and Washington state freshwater sponges. The chemical potential of these strains has been captured in a fraction library at 10 mg/mL in DMSO for evaluation of biological activity. To evaluate the entire collection for biological activity, we have not been able to chase down the last few strains due to impurities or missing cryovials. Over the last three years, the Carlson Lab Actinomycete Collection has provided research opportunities for numerous undergraduate researchers. Due to repeated access to cryovials by multiple researchers, the purity of some vials may have been compromised. To determine the purity of our cryopreserved microbes, a large-scale purity check operation took place. All strains in our collection were plated onto A1 or A1Di media - starch, yeast, peptone, and agar with or without salt water, respectively. After a few weeks of growth on salad media, the purity of each cryovial was evaluated. Contaminated cryovials will be removed from the collection and roster of pure strains. Any actinomycetes that can be isolated from the contaminated plates will be grown in liquid culture, cryopreserved anew, and given the next available record number. Pure actinomycetes were photographed to prepare a visual reference gallery, providing a standardized guide to ensure consistency in future work. The progress of cultivation to fractionation and purity check will be presented.
Actinomycete library preparation and large scale cultivation
Actinomycetes are a phylum of gram-positive bacteria known to dedicate a large portion of their genome to secondary metabolite production. These secondary metabolites are small, drug-like molecules that have potent biological activity useful in inspiring new drug candidates. The Carlson Lab Actinomycete Collection is composed of approximately 100 strains. These have been isolated from the Great Lakes freshwater sponges, Puerto Rico sediments, and Washington state freshwater sponges. The chemical potential of these strains has been captured in a fraction library at 10 mg/mL in DMSO for evaluation of biological activity. To evaluate the entire collection for biological activity, we have not been able to chase down the last few strains due to impurities or missing cryovials. Over the last three years, the Carlson Lab Actinomycete Collection has provided research opportunities for numerous undergraduate researchers. Due to repeated access to cryovials by multiple researchers, the purity of some vials may have been compromised. To determine the purity of our cryopreserved microbes, a large-scale purity check operation took place. All strains in our collection were plated onto A1 or A1Di media - starch, yeast, peptone, and agar with or without salt water, respectively. After a few weeks of growth on salad media, the purity of each cryovial was evaluated. Contaminated cryovials will be removed from the collection and roster of pure strains. Any actinomycetes that can be isolated from the contaminated plates will be grown in liquid culture, cryopreserved anew, and given the next available record number. Pure actinomycetes were photographed to prepare a visual reference gallery, providing a standardized guide to ensure consistency in future work. The progress of cultivation to fractionation and purity check will be presented.
