Penn State University 2006:Growthmedia: Difference between revisions

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=Media=
=Media=


LB MEDIA
===<font color="red">LB Media</font>===
(Per L)
(Per L)
10 g bactotryptone
*10 g bactotryptone
5 g yeast extract
*5 g yeast extract
5 g NaCl
*5 g NaCl
pH to 7.0  
*pH to 7.0  


M9 MINIMAL MEDIA (SUPPLEMENTED W/ 0.1% YE)
===<font color="red">M9 Minimal Media</font>===
(SUPPLEMENTED W/ 0.1% YE)
Final concentrations:
Final concentrations:
1X M9 salts (autoclaved)
*1X M9 salts (autoclaved)
1 mM MgSO4 (autoclaved)
*1 mM MgSO4 (autoclaved)
0.16% glycerol (autoclaved)
*0.16% glycerol (autoclaved)
0.148 μM thiamine/vitamin B1 (filter sterilized)
*0.148 μM thiamine/vitamin B1 (filter sterilized)
0.1% casamino acids (filter sterilized)
*0.1% casamino acids (filter sterilized)
0.1% Yeast Extract (autoclaved)
*0.1% Yeast Extract (autoclaved)
 
===<font color="green">Procedure:</font>===
Procedure:
1. These solutions should already be made,  
these solutions should already be made,  
so add together w/sterile technique:
so add together w/sterile technique:


200 mL 5X M9 salts
*200 mL 5X M9 salts
1 mL 1 M MgSO4
*1 mL 1 M MgSO4
2.27 mL 70% glycerol
*2.27 mL 70% glycerol
50 μL thiamine (10mg/mL)  
*50 μL thiamine (10mg/mL)  
5 mL casamino acids (200 mg/mL)
*5 mL casamino acids (200 mg/mL)
10 mL 0.1g/mL Yeast Extract  
*10 mL 0.1g/mL Yeast Extract  
782 mL sterile dH2O                          .
*782 mL sterile dH2O                          .
1 L M9 media (for plates just add 15 g bacto agar to recipe)
*1 L M9 media (for plates just add 15 g bacto agar to recipe)


SOB MEDIA
===<font color="red">SOB Media</font>===
Final concentrations
Final concentrations
0.5% Yeast Extract
*0.5% Yeast Extract
2% Tryptone
*2% Tryptone
10mM NaCl
*10mM NaCl
2.5 mM KCl
*2.5 mM KCl
10 mM MgCl2
*10 mM MgCl2
10 mM MgSO4
*10 mM MgSO4


Procedure:
===<font color="green">Procedure</font>===
To 1 L, add:  
1. To 1 L, add:  
20 g bactotryptone
*20 g bactotryptone
5 g bacto-yeast extract
*5 g bacto-yeast extract
0.5 g NaCl
*0.5 g NaCl
0.19 g KCl
*0.19 g KCl
Adjust to pH 7.0 w/ NaOH
*Adjust to pH 7.0 w/ NaOH


Autoclave on slow exhaust for 25 min.
2. Autoclave on slow exhaust for 25 min.


Add filter-sterilized MgCl2, MgSO4 solution to give final Mg2+ conc. of 20 mM (i.e. 10 mM MgCl2, 10 mM MgSO4)
3. Add filter-sterilized MgCl2, MgSO4 solution to give final Mg2+ conc. of 20 mM (i.e. 10 mM MgCl2, 10 mM MgSO4)


SOC MEDIA
SOC MEDIA

Revision as of 16:59, 24 October 2006

Media

LB Media

(Per L)

  • 10 g bactotryptone
  • 5 g yeast extract
  • 5 g NaCl
  • pH to 7.0

M9 Minimal Media

(SUPPLEMENTED W/ 0.1% YE) Final concentrations:

  • 1X M9 salts (autoclaved)
  • 1 mM MgSO4 (autoclaved)
  • 0.16% glycerol (autoclaved)
  • 0.148 μM thiamine/vitamin B1 (filter sterilized)
  • 0.1% casamino acids (filter sterilized)
  • 0.1% Yeast Extract (autoclaved)

Procedure:

1. These solutions should already be made, so add together w/sterile technique:

  • 200 mL 5X M9 salts
  • 1 mL 1 M MgSO4
  • 2.27 mL 70% glycerol
  • 50 μL thiamine (10mg/mL)
  • 5 mL casamino acids (200 mg/mL)
  • 10 mL 0.1g/mL Yeast Extract
  • 782 mL sterile dH2O .
  • 1 L M9 media (for plates just add 15 g bacto agar to recipe)

SOB Media

Final concentrations

  • 0.5% Yeast Extract
  • 2% Tryptone
  • 10mM NaCl
  • 2.5 mM KCl
  • 10 mM MgCl2
  • 10 mM MgSO4

Procedure

1. To 1 L, add:

  • 20 g bactotryptone
  • 5 g bacto-yeast extract
  • 0.5 g NaCl
  • 0.19 g KCl
  • Adjust to pH 7.0 w/ NaOH

2. Autoclave on slow exhaust for 25 min.

3. Add filter-sterilized MgCl2, MgSO4 solution to give final Mg2+ conc. of 20 mM (i.e. 10 mM MgCl2, 10 mM MgSO4)

SOC MEDIA SOB + 20 mM (final) glucose (aka dextrose)


Main

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