SMM - Revision history

MFeeney: /* Preparation */

2022-07-19T10:25:09Z

Preparation

← Older revision		Revision as of 10:25, 19 July 2022
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	*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)		*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
	*2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final		*2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final
	*10 mL TES ~~bugger~~ (0.25M, pH 7.2) - 25mM final		*10 mL TES buffer (0.25M, pH 7.2) - 25mM final
	*1 mL NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final		*1 mL NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final
	*2 mL glucose (50% w/v)		*2 mL glucose (50% w/v)

Tommclean: /* SMM (Supplemented liquid minimal media) */

2019-08-01T15:52:02Z

SMM (Supplemented liquid minimal media)

← Older revision		Revision as of 15:52, 1 August 2019
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	=SMM (Supplemented liquid minimal media)=		=SMM (Supplemented liquid minimal media)=
	~~Strauch et al~~.~~, 1991<sup>1</sup>~~		SMM is a supplemented minimal medium suitable for growth of many ''Streptomyces'' sp.

	==Preparation==		==Preparation==
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	#With the exception of the trace element solution each solution must be made up and autoclaved separately		#With the exception of the trace element solution each solution must be made up and autoclaved separately
	#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water		#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water

			==References==
			Strauch E, Takano E, Baylts H, Bibb M. The stringent response in ''Streptomyces coelicolor'' A3(2). ''Molecular Microbiology''. 1991;5(2):289-298.

Tommclean at 15:42, 1 August 2019

2019-08-01T15:42:07Z

← Older revision		Revision as of 15:42, 1 August 2019
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	*2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final		*2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final
	*10 mL TES bugger (0.25M, pH 7.2) - 25mM final		*10 mL TES bugger (0.25M, pH 7.2) - 25mM final
	*~~1mL~~ NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final		*1 mL NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final
	*2 mL glucose (50% w/v)		*2 mL glucose (50% w/v)
	*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)		*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
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	*100 mL Total		*100 mL Total
	*25 ml/L Glycine (0.5% final)		*25 ml/L Glycine (0.5% final)

			Trace element solution:
			0.1 g/L of the following:
			*ZnSO<sub>4</sub>.7H<sub>2</sub>O
			*FeSO<sub>4</sub>.7H<sub>2</sub>O
			*MnCl<sub>2</sub>.4H<sub>2</sub>O
			*CaCl<sub>2</sub>.6H<sub>2</sub>O
			*NaCl
			Make fresh every 2-4 weeks and filter-sterilise. Store at 4º


	'''Instructions'''		'''Instructions'''
	#~~Dissolve all~~ of the ~~ingredients except the agar~~ and ~~the glucose in the distilled water~~		#With the exception of the trace element solution each solution must be made up and autoclaved separately
	#~~Adjust~~ the pH to ~~7.0-7.2~~		#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water
	~~#Dispense 200ml into flasks each containing 2g agar~~
	~~#Autoclave~~
	~~#After autoclaving, add 4 ml~~ sterile ~~50% glucose solution (autoclaved separately or filter-sterilized) to each flask~~
	~~#Add any other growth factors required for auxotrophic mutants~~

Tommclean: Created page with "=SMM (Supplemented liquid minimal media)= Strauch et al., 1991¹ ==Preparation== Per litre: 81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) 2.5 mL Mg_{2
2019-08-01T15:30:50Z
Created page with "=SMM (Supplemented liquid minimal media)= Strauch et al., 19911 ==Preparation== Per litre: 81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) 2.5 mL Mg2</su..."
New page
=SMM (Supplemented liquid minimal media)=

Strauch et al., 19911

==Preparation==

Per litre:

81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)

2.5 mL Mg2SO4.7H2O - 5mM final

10 mL TES bugger (0.25M, pH 7.2) - 25mM final

1mL NaH2PO4 + K2HPO4 (50mM each) - 1mM final

2 mL glucose (50% w/v)

1 mL Antifoam 289 (Sigma A 5551)(1% w/v)

0.1 mL Trace element solution

1 mL Difco Casaminoacids (20% w/v)

100 mL Total

25 ml/L Glycine (0.5% final)

'''Instructions'''

#Dissolve all of the ingredients except the agar and the glucose in the distilled water

#Adjust the pH to 7.0-7.2

#Dispense 200ml into flasks each containing 2g agar

#Autoclave

#After autoclaving, add 4 ml sterile 50% glucose solution (autoclaved separately or filter-sterilized) to each flask

#Add any other growth factors required for auxotrophic mutants}