AB medium: Difference between revisions
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#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); | |||
# 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock) | # 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock) | ||
# 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>.) | # 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>.) | ||
Revision as of 02:16, 31 March 2008
Purpose
- 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 [1].
Recipe
| for 1 liter AB medium |
|---|
| 2 g (NH4)2SO4 |
| 6 g Na2HPO4 |
| 3 g KH2PO4 |
| 3 g NaCl |
| 200 ml H2O |
Prepare and autoclave.
| 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!
Different things can or should be added:
- Carbon source (has to be added):
- 0.2% glucose (10ml of 20% or 4ml of 50% sterile stock)
- 0.4% acetate (20ml of 20% sterile sodium-acetate stock)
- Amino acids (check AA requirement of your strain):
- 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);
- 10μg/ml thiamine (1 ml of sterile 10 mg/ml stock)
- 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 [2].)
References
-
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
- Jensen KF. The Escherichia coli K-12 "wild types" W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol. 1993 Jun;175(11):3401-7. DOI:10.1128/jb.175.11.3401-3407.1993 |
Contact
- Please feel free to edit this page, add comments on the talk page or contact the page beginner Torsten Waldminghaus.
