|
Components
|
Stock solution (mg/ml)
|
Vol stock solution (ml) for 1 liter
|
Final conc (mmol/l) of 1xCDB
|
|---|
|
Buffer solution (10X)
| |
100.0
| |
|
K2HPO4
|
34.8
| |
20.0
|
|
KH2PO4
|
27.2
| |
20.0
|
|
Salt solution (20X)
| |
50.0
| |
|
NaCl
|
116.0
| |
100.00
|
|
K2SO4
|
20.0
| |
5.74
|
|
MgCl2, 6 H2O
|
8.2
| |
2.02
|
|
NH4Cl
|
4.4
| |
4.11
|
|
EDTA
|
0.074
| |
0.01
|
|
Amino acid mix 1 (100X)
| |
10.0
| |
|
L-Arginine HCl
|
15.0
| |
0.71
|
|
L-serine
|
5.0
| |
0.48
|
|
L-leucine
|
9.0
| |
0.69
|
|
L-isoleucine
|
3.0
| |
0.23
|
|
L-valine
|
6.0
| |
0.51
|
|
L-proline
|
5.0
| |
0.43
|
|
L-phenylalanine
|
5.0
| |
0.30
|
|
L-alanine
|
10.0
| |
1.12
|
|
L-histidine
|
5.0
| |
0.32
|
|
L-threonine
|
5.0
| |
0.42
|
|
L-lysine
|
5.0
| |
0.30
|
|
L-glycine
|
2.5
| |
0.33
|
|
L-trypthophan
|
8.0
| |
0.39
|
|
Amino acid mix 2 (10X)
| |
100.0
| |
|
L-aspartate
|
5.0
| |
3.76
|
|
L-glutamate
|
13.0
| |
8.83
|
|
Individual amino acids
| | | |
|
L-cysteine/HCl †
|
17.5
|
3.0
|
0.35
|
|
L-cysteine †
|
12.0
|
3.5
|
0.15
|
|
L-methionine
|
14.9
|
1.0
|
0.10
|
|
Vitamin mix (50X)
| |
0.2
| |
|
NAD
|
10.0
| |
0.0030
|
|
Thiamine HCl (Vitamine B1)
|
10.0
| |
0.0060
|
|
Calcium pantothenate (Vitamine B5)
|
10.0
| |
0.0040
|
|
Individual components
| | | |
|
Oxaloacetate, 2 H2O (10X)
|
2.5
|
100.0
| |
|
NaHCO3 (2000×)
|
84.0
|
0.5
|
1.49
|
|
Biotin (Vitamine H) (500X)
|
0.25
|
2.0
|
0.50
|
|
Fe(NO3)3, 9 H2O (1000X)
|
4.0
|
1.0
|
0.0021
|
|
CaCl2, 2 H2O (1000X)
|
37.0
| |
0.01
|
| | | |
0.25
|
|
Total (ml)
| |
372.2
| |
|
MilliQ water (ml) *
| |
627.8
| |
- The broth was prepared as described in the text.
- *: To make portions of 500 ml 2 × CDB without methionine and cysteine, only 127.8 ml of MilliQ water was added. The batches were sterilized by filtration, aliquoted, and stored at −20°C.
- †: Cysteine was prepared freshly and dissolved in 1 M HCl prior to each experiment.
