403 .
BSK-MEDIUM
What is this page?
Equipment needed:
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 1000 | ml | ||
|
|
Solution B | 200 | ml | ||
|
|
Solution C | 143 | ml | ||
|
|
Solution D | 86 | ml | ||
| 1 Combine the warm (37°C) solutions A, B, C, D, and filter-sterilize the warm medium. Solution B can also be added after filtration. | |||||
| 2 Alternatively, buy supplemented or un-supplemented BSK-H medium (Fa. Bio&SELL or Sigma) and supplement the medium according to manufacturer's advice. Sterilize the mixed and warm (37°C) medium by filtration. | |||||
Solution A #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
CMRL 1066
(GIBCO 042-1545, glutamin 10 x) |
100.0 | ml | ||
|
|
Proteose peptone no. 2
(DIFCO 0121-01-3) |
5.0 | g | ||
|
|
Tryptone peptone
(DIFCO 0123-01) |
1.0 | g | ||
|
|
Proteose yeastolate
(TC) (DIFCO 5577-15) |
1.0 | g | ||
|
|
HEPES buffer
(acid) (SIGMA H3375) |
6.0 | g | ||
|
|
Sodium citrate | 0.7 | g | ||
|
|
Glucose
(alpha-, D+) |
3.0 | g | ||
|
|
Na-pyruvate | 0.8 | g | ||
|
|
N-Acetylglucosamine | 0.4 | g | ||
|
|
MgCl2 x 6 H2O | 0.3 | g | ||
|
|
L-Glutamine | 0.1 | g | ||
|
|
Double distilled water
(filter-sterilized) |
900.0 | ml | ||
| 1 Stir slowly at 4 - 10°C for 3 hours, adjust pH to 7.6 with 5 M NaOH. Filter sterilize. | |||||
Solution B #
200
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Gelatine | 14 | g | ||
|
|
Double distilled water | 200 | ml | ||
| 1 Dissolve 14.0 g gelatine in 200 ml bidest. water, autoclave 15 minutes at 115°C. | |||||
Solution C #
143
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Bovine serum albumin
(Sigma No A9647) |
50.05 | g | ||
|
|
Double distilled water | 143.00 | ml | ||
| 1 Dissolve 50.05 g bovine serum albumine, fract. V (important: Sigma No A9647) in 143 ml bidest. water, or use BSA-solution Sigma A7409. | |||||