NMMP: Difference between revisions
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#Make up the phosphate buffer solution, carbon source, and any other required growth factors, and autoclave these separately | #Make up the phosphate buffer solution, carbon source, and any other required growth factors, and autoclave these separately | ||
#Just before using, add: | #Just before using, add: | ||
*15 mL NaH<sub>2</sub>PO<sub>4</sub>/K<sub>2</sub>HPO<sub>4</sub> buffer (0.1M, pH 6.8) | **15 mL NaH<sub>2</sub>PO<sub>4</sub>/K<sub>2</sub>HPO<sub>4</sub> buffer (0.1M, pH 6.8) | ||
*2.5 mL carbon source (20%) | **2.5 mL carbon source (20%) | ||
*2.5 mL any other required growth factors (add 2.5 mL sterile water if none) | **2.5 mL any other required growth factors (add 2.5 mL sterile water if none) | ||
==Uses== | ==Uses== | ||
Revision as of 10:08, 29 July 2019
Minimal liquid medium (NMMP)
NMMP medium is used to obtain dispersed growth of Streptomyces sp. with specific carbon sources.
Preparation
Make up the following solutions:
Minor elements solution: (per litre)
- 1g ZnSO4*7H2O
- 1g FeSO4*7H2O
- 1g MnCl2*4H2O
- 1g CaCl2, anhydrous
Phosphate buffer
- Mix 0.1M solutions of NaH2PO4 and K2HPO4 to give pH 6.8
Carbon source
- 20% solution, sterilized separately
NMMP
- 2 g (NH4)2SO4
- 5 g Difco Casaminoacids
- 0.6g MgSO4*7H2O
- 50g PEG 6000
- 1 mL minor elements solution
- 800 mL ddH2O
Instructions
- Make up the NMMP solution described above
- Dispense in 80 mL aliquots
- Autoclave
- Make up the phosphate buffer solution, carbon source, and any other required growth factors, and autoclave these separately
- Just before using, add:
- 15 mL NaH2PO4/K2HPO4 buffer (0.1M, pH 6.8)
- 2.5 mL carbon source (20%)
- 2.5 mL any other required growth factors (add 2.5 mL sterile water if none)
Uses
- Routine growth of Streptomyces sp. in liquid culture
Notes
References
Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK
