反应详情

温度

55 °C

Acetamide (Aldrich 99% sublimed)—0.6 g/L; CsCl—1.68 g/L; Glucose—20 g/L; KH2PO4—20 g/L; MgSO4.7H2O—0.6 g/L; CaCl2.2H2O—0.6 g/L; 1000× salts (see below)—1 ml. pH adjusted to 5.5 and volume brought to 300 ml. Filter sterilized with 0.22 micron filter and warmed to 55° C. in an oven. To 700 ml water Noble Agar (low-melt for top agar) 20 g and Sorbitol 218 g was added and then autoclaved. This mixture was cooled to 55° C., and filter sterilized, acetamide mix was added. Plates or tubes were poured. 1000× Salts—FeSO4.7H2O (0.5 g/100 ml); MnSO4.H2O (0.16 g/100 ml); ZnSO4.7H2O (0.14 g/100 ml); CoCl2.6H2O (0.1 g/100 ml) and filter sterilize with 0.22 micron filter. Potato Dextrose Agar (PDA Difco Dehydrated Culture Media)—Potatoes, infusion from 200 g/L; Dextrose, 20 g/L and Agar, 15 g/L were mixed well in 50-80% final volume of dH2O, and then brought to 100% final volume. This mixture was autoclaved, cooled to 55° C. and poured. To make up 1% skim milk agar for a pH 3.5 media PDA was prepared as above and to 100 ml molten PDA, 1.8 ml 10% tartaric acid and 12.5 ml sterilized 8% skim milk was added and plates were poured. To pre-sterilize skim milk, 8% skim milk (Difco) was autoclaved for 10 minutes, 122-123° C., and chamber pressure during exposure of 32-35 psi. The mixture was removed, cooled and stored at room temperature.

1. filtrationFilter
2. filtrationfilter
3. additionTo 700 ml water Noble Agar (low-melt for top agar) 20 g and Sorbitol 218 g was added
4. temperatureThis mixture was cooled to 55° C.
5. filtrationfilter
6. additionacetamide mix
7. additionwas added
8. additionPlates or tubes were poured
9. filtrationCoCl2.6H2O (0.1 g/100 ml) and filter sterilize with 0.22 micron
10. filtrationfilter
11. additionDextrose, 20 g/L and Agar, 15 g/L were mixed well in 50-80% final volume of dH2O
12. temperaturecooled to 55° C.
13. additionpoured
14. customwas prepared as above and to 100 ml molten PDA, 1.8 ml 10% tartaric acid and 12.5 ml
15. additionwas added
16. additionplates were poured
17. customThe mixture was removed
18. temperaturecooled
19. customstored at room temperature