1825 .
MDM
What is this page?
Equipment needed:
Autoclave
Filter
Final pH:
6.0 - 7.0
Description:
Minimal medium for Clostridioides difficile. It is a minimal medium with only required amino acids, growth can be strongly enhanced by the addition of L-threonine. Adapted from Karlsson S, Burman LG, Akerlund T. Suppression of toxin production in Clostridium difficile VPI 10463 by amino acids. Microbiology. 1999;145(Pt 7):1683–93.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Salt mix 10x | 100 | ml | ||
|
|
Amino acids mix 5x | 200 | ml | ||
|
|
Trace salt mix 100x | 10 | ml | ||
|
|
Iron stock solution 2.0 g/L | 3 | ml | ||
|
|
Vitamine mixture | 5 | ml | ||
|
|
Glucose stock 20% | 10 | ml | ||
| 1 Make up to 1000 ml with sterile distilled water. | |||||
| 2 anaerobize with nitrogen, can be stored anaerobically for a limited time | |||||
Salt mix 10x #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 | 50 | g/l | ||
|
|
NaH2PO4 x H2O | 20 | g/l | ||
|
|
KH2PO4 | 9 | g/l | ||
|
|
NaCl | 9 | g/l | ||
| 1 autoclave | |||||
Trace salt mix 100x #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 4.000 | g/l | ||
|
|
CaCl2 x 2 H2O | 2.600 | g/l | ||
|
|
MgCl2 x 6 H2O | 2.000 | g/l | ||
|
|
MnCl2 x 4 H2O | 1.000 | g/l | ||
|
|
CoCl2 x 6 H2O | 0.100 | g/l | ||
|
|
NaHSeO3 , almost no effect on growth in CDMM, more important in MDM | 0.015 | g/l | ||
| 1 steril filtration | |||||
Iron stock solution 2.0 g/L #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
FeSO4 x 7 H2O
(Sigma Aldrich!) |
2 | g/l | ||
| 1 steril filtration, store in aliquots at -20°C | |||||
Vitamine mixture #
500
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
D-Biotin | 0.06 | g/l | ||
|
|
Calcium D-(+)-pantothenate | 0.20 | g/l | ||
|
|
Pyridoxine | 0.20 | g/l | ||
| 1 steril filtration, store aliquots at -20°C | |||||
Glucose stock 20% #
500
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
alpha-D-Glucose , not required but growth enhancing | 200 | g/l | ||
| 1 steril filtration | |||||
Amino acids mix 5x #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
L-Cysteine | 10.0 | g | ||
|
|
L-Tryptophan | 0.5 | g | ||
|
|
L-Leucine | 2.0 | g | ||
|
|
L-Valine | 1.5 | g | ||
|
|
L-Proline | 10.0 | g | ||
|
|
L-Isoleucine | 1.5 | g | ||
|
|
L-Methionine | 1.0 | g | ||
| 1 Adjust pH to 6.0-7.0. | |||||
| 2 Filtrate. | |||||
| 3 Store at -20°C | |||||