SPY: Difference between revisions
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==References== | ==References== | ||
[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4421039/ Imai, Y., Sato, S., Tanaka, Y., Ochi, K. and Hosaka, T. (2015). Lincomycin at Subinhibitory Concentrations Potentiates Secondary Metabolite Production by Streptomyces spp. Applied and Environmental Microbiology, 81(11), pp.3869-3879.] |
Latest revision as of 15:56, 1 August 2019
Salt Peptone Yeast Agar (SPY)
Salt Peptone Yeast agar is a metal-rich media used to stimulate the activation of cryptic biosynthetic gene clusters as part of antibiotic discovery.
Preparation
Per Litre:
- 30g Soluable Starch
- 15g pancreatic digest of casein
- 15g peptic digest of animal tissue
- 2g Yeast Extract
- 10g MgSO4
- 50mg CaCl2
- 30mg ZnSO4
- 25mg FeSO4
- 16mg CuSO4
- 12mg MnSO4
- 20g agar
- NaOH (to adust pH)
- 1L dH2O
Instructions
- On a heated stiring block, warm the water and add the pancreatice digest of casin, the peptic digest of animal tissue, the yeast extract and the trace elements.
- Add 2g of agar into each of 10 250mL conical flasks.
- Use NaOH to adjust the pH of the solution to pH 7.2.
- Dissolve the soluable starch in the solution. It can take a while for the starch to dissolve.
- Pour 100mL of solution into each flask.
- Autoclave
Uses
Notes
- To dissolve the soluable starch, it is often required to make the media on a hot plate. Do not allow the solution to boil.
- To add the trace elements, it may be easier to make concentrated stock solution which is then added to the media prior to pH adjustment.