AB medium

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(Recipe)
 
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==Purpose==
==Purpose==
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*AB medium is a minimal growth medium used for bacterial cultures. It has some changes compared to [[M9 medium]] and is supposed to course less problems with precipitation <cite>Clark-JMB-1967</cite>.
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*AB medium is a minimal growth medium used for bacterial cultures. It has some changes compared to [[M9 medium]] and is supposed to cause less problems with precipitation <cite>Clark-JMB-1967</cite>.
==Recipe==
==Recipe==
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Mix 200ml 5xA and 800ml B to get 1 liter AB. Do never add B salts to 5xA!
Mix 200ml 5xA and 800ml B to get 1 liter AB. Do never add B salts to 5xA!
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 +
If you want''' agar-plates''' add 16g agar to 1xB, autoclave and than add 5xA.
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#Carbon source (has to be added):  
#Carbon source (has to be added):  
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## 0.2% glucose (10ml of 20% or 4ml of 50% sterile stock)  
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## 0.2% [[glucose]] (10ml of 20% or 4ml of 50% sterile stock)  
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## 0.4% acetate (20ml of 20% sterile sodium-acetate stock)  
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## 0.4% acetate (20ml of 20% sterile [[sodium-acetate]] stock)  
#Amino acids (check AA requirement of your [[E. coli genotypes | strain]]):  
#Amino acids (check AA requirement of your [[E. coli genotypes | strain]]):  
## 0.5% [[casamino acids]] (50ml of 10% sterile stock)  
## 0.5% [[casamino acids]] (50ml of 10% sterile stock)  
## For CM735 add 20 μg/ml methionine (10 ml of 2mg/ml stock), 20 μg/ml tryptophane (5 ml of 4mg/ml stock) and 25 μg/ml histidine (1 ml of 25 mg/ml stock);  
## For CM735 add 20 μg/ml methionine (10 ml of 2mg/ml stock), 20 μg/ml tryptophane (5 ml of 4mg/ml stock) and 25 μg/ml histidine (1 ml of 25 mg/ml stock);  
# 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock)
# 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock)
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# 25μg/ml uridine (0.25 ml of sterile 10 mg/ml stock)(uridine makes E. coli K12 wildtypes as MG1655 of W3110 grow better because they are starving from pyrimidine <cite>Jensen-JB-1993</cite>.)
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# 25μg/ml uridine (2.5 ml of sterile 10 mg/ml stock)(uridine makes E. coli K12 wildtypes as MG1655 of W3110 grow better because they are starving from pyrimidine <cite>Jensen-JB-1993</cite>.)
==References==
==References==
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*Please feel free to edit this page, add comments on the talk page or contact the page beginner [[User:Torsten Waldminghaus|Torsten Waldminghaus]].  
*Please feel free to edit this page, add comments on the talk page or contact the page beginner [[User:Torsten Waldminghaus|Torsten Waldminghaus]].  
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[[Category:Material]][[Category:Defined Media]][[Category:Media]]
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[[Category:Material]][[Category:Defined Media]]

Current revision

Contents

Purpose

  • AB medium is a minimal growth medium used for bacterial cultures. It has some changes compared to M9 medium and is supposed to cause less problems with precipitation [1].

Recipe

5xA
for 1 liter AB medium
2 g (NH4)2SO4
6 g Na2HPO4
3 g KH2PO4
3 g NaCl
200 ml H2O

Prepare and autoclave.


1xB
for 1 liter AB medium
1 ml 0.1 M CaCl2 (sterile)
1 ml 1.0 M MgCl2 (sterile)
1 ml 0.003 M FeCl3 (sterile)
797 ml H2O (sterile)

Mix 200ml 5xA and 800ml B to get 1 liter AB. Do never add B salts to 5xA!

If you want agar-plates add 16g agar to 1xB, autoclave and than add 5xA.


Different things can or should be added:

  1. Carbon source (has to be added):
    1. 0.2% glucose (10ml of 20% or 4ml of 50% sterile stock)
    2. 0.4% acetate (20ml of 20% sterile sodium-acetate stock)
  2. Amino acids (check AA requirement of your strain):
    1. 0.5% casamino acids (50ml of 10% sterile stock)
    2. For CM735 add 20 μg/ml methionine (10 ml of 2mg/ml stock), 20 μg/ml tryptophane (5 ml of 4mg/ml stock) and 25 μg/ml histidine (1 ml of 25 mg/ml stock);
  3. 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock)
  4. 25μg/ml uridine (2.5 ml of sterile 10 mg/ml stock)(uridine makes E. coli K12 wildtypes as MG1655 of W3110 grow better because they are starving from pyrimidine [2].)

References

  1. Clark, DJ and Maaløe, O DNA replication and the division cycle in Escherichia coli Journal of Molecular Biology Volume 23, Issue 1, 14 January 1967, Pages 99-112 abstract [Clark-JMB-1967]
  2. Jensen KF. . pmid:8501045. PubMed HubMed [Jensen-JB-1993]

Contact

  • Please feel free to edit this page, add comments on the talk page or contact the page beginner Torsten Waldminghaus.
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