NMMP - Revision history

MFeeney at 10:26, 29 July 2019

2019-07-29T10:26:11Z

← Older revision		Revision as of 10:26, 29 July 2019
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	*Agar can be added (20 g/L) to make solid NMMP.<sup>[3]</sup>		*Agar can be added (20 g/L) to make solid NMMP.<sup>[3]</sup>

	*A modified version of NMMP suitable for <sup>14</~~sub~~>N/<sup>15</sup>N labelling has been developed.<sup>[4]</sup>		*A modified version of NMMP suitable for <sup>14</sup>N/<sup>15</sup>N labelling has been developed.<sup>[4]</sup>

MFeeney at 10:25, 29 July 2019

2019-07-29T10:25:24Z

← Older revision		Revision as of 10:25, 29 July 2019
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	#Autoclave		#Autoclave
	#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:
	15 mL NaH<sub>2</sub>PO<sub>4</sub>/K<sub>2</sub>HPO<sub>4</sub> buffer (0.1M, pH 6.8)		Just before using, add:
	2.5 mL carbon source (20%)		*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 any other required growth factors (add 2.5 mL sterile water if none)		*2.5 mL carbon source (20%)
			*2.5 mL any other required growth factors (add 2.5 mL sterile water if none)

	==Uses==		==Uses==

MFeeney at 10:23, 29 July 2019

2019-07-29T10:23:53Z

← Older revision		Revision as of 10:23, 29 July 2019
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	=Minimal liquid medium (NMMP)=		=Minimal liquid medium (NMMP)=

	NMMP medium is used to obtain dispersed growth of ''Streptomyces'' sp. with specific carbon sources.		NMMP medium is used to obtain dispersed growth of ''Streptomyces'' sp. with specific carbon sources.<sup>[2]</sup>

	==Preparation==		==Preparation==
	Make up the following solutions:		Make up the following solutions:

	Minor elements solution: (per litre)		'''Minor elements solution''': (per litre)
	1g ZnSO<sub>4</sub>7H<sub>2</sub>O		1g ZnSO<sub>4</sub>7H<sub>2</sub>O
	1g FeSO<sub>4</sub>7H<sub>2</sub>O		1g FeSO<sub>4</sub>7H<sub>2</sub>O
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	*1g CaCl<sub>2</sub>, anhydrous		*1g CaCl<sub>2</sub>, anhydrous

	Phosphate buffer		'''Phosphate buffer'''
	*Mix 0.1M solutions of NaH<sub>2</sub>PO<sub>4</sub> and K<sub>2</sub>HPO<sub>4</sub> to give pH 6.8		*Mix 0.1M solutions of NaH<sub>2</sub>PO<sub>4</sub> and K<sub>2</sub>HPO<sub>4</sub> to give pH 6.8

	Carbon source		'''Carbon source'''
	*20% solution, sterilized separately		*20% solution, sterilized separately

	NMMP		'''NMMP'''
	*2 g (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>		*2 g (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>
	*5 g Difco Casaminoacids		*5 g Difco Casaminoacids
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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==
Line 41:		Line 41:

	==Notes==		==Notes==
			*Using PEG-6000 and adding stainless steel springs in the culture flasks allows for good dispersed growth of many ''Streptomyces'' strains.<sup>[1]</sup>

			*Agar can be added (20 g/L) to make solid NMMP.<sup>[3]</sup>

			*A modified version of NMMP suitable for <sup>14</sub>N/<sup>15</sup>N labelling has been developed.<sup>[4]</sup>


	==References==		==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		[1] Hodgson (1982). Glucose Repression of Carbon Source Uptake and Metabolism in ''Streptomyces coelicolor'' A3(2) and its Perturbation in Mutants Resistant to 2-Deoxyglucose. Journal of General Microbiology 128: 2417-2430.

			[2] 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

			[3] Rico, S., Yepes, A., Rodríguez, H., Santamaría, J., Antoraz, S., Krause, E. M., … Santamaría, R. I. (2014). Regulation of the AbrA1/A2 two-component system in ''Streptomyces coelicolor'' and the potential of its deletion strain as a heterologous host for antibiotic production. PloS one, 9(10), e109844. doi:10.1371/journal.pone.0109844

			[4] Świątek, M. A., Gubbens, J., Bucca, G., Song, E., Yang, Y. H., Laing, E., … van Wezel, G. P. (2013). The ROK family regulator Rok7B7 pleiotropically affects xylose utilization, carbon catabolite repression, and antibiotic production in ''Streptomyces coelicolor''. Journal of bacteriology, 195(6), 1236–1248. doi:10.1128/JB.02191-12

MFeeney at 10:08, 29 July 2019

2019-07-29T10:08:20Z

← Older revision		Revision as of 10:08, 29 July 2019
Line 32:		Line 32:
	#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==

MFeeney: Created page with "=Minimal liquid medium (NMMP)= NMMP medium is used to obtain dispersed growth of ''Streptomyces'' sp. with specific carbon sources. ==Preparation== Make up the following sol..."

2019-07-29T10:07:30Z

Created page with "=Minimal liquid medium (NMMP)= NMMP medium is used to obtain dispersed growth of ''Streptomyces'' sp. with specific carbon sources. ==Preparation== Make up the following sol..."

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=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