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Mycoplasma Broth Recipes－PPLO Broth
关键词： Mycoplasma Broth Recipes－PPLO Broth来源： 互联网
| 200 ml | 1000 ml | 4000 ml | |
| Mycoplasma broth base (Gibco) | 4.5 | 22.5 | 90.0 |
| DNA (Sigma, Degraded free acid, type IV) | 0.05 | 0.20 | 0.80 |
| Glass Distilled Water | 142 ml | 710 ml | 2840 ml |
| Phenol Red (1% aqueous) | 0.4 ml | 2.0 ml | 8.0 ml |
| For Agar Media using Noble Agar (Difco) | 2.0 | 10.0 | 40.0 |
| Using Seaplaque Agarose (FMC corp. Marine Colloids Div.) | 1.6 | 8.0 | 32.0 |
Adjust pH to 7.8 - 8.0 with 1N NaOH
Autoclave at 121°C for 15 min. at 15 lbs pressure, slow exhaust.
Allow to cool for liquid medium, and bring to 50°C in a water bath for agar
Add aseptically:
| 200 ml | 1000ml | 4000 ml | |
| Agamma horse serum | 40 ml | 200 ml | 800 ml |
| 25% Yeast Extract | 10ml | 50 ml | 200 ml |
| Penicillin G (100,000 units/ml) | 1 ml | 5 ml | 20 ml |
| Cefobid 2.5 mg/ml (50:50 EtOH:50 water) | 2 ml | 10 ml | 40 ml |
| 50 % Dextrose (filter sterilized) | 2 ml | 10 ml | 40 ml |
pH should be around 7.8-8.0
Store at 4°C for one month or freeze for 3 months
Aseptically dispense agar into petri dishes with a minimum depth of 4mm, store in sealed plastic bags at 4°C for 3 weeks.
Mycoplasma Broth
| 4 L | 1 L | 0.5 L | |
| Dist. Water | 2640 ml | 660 ml | 330 ml |
| PPLO Broth Base | 84 g | 21 g | 10.5 g |
| Glucose | 20 g | 5 g | 2.5 g |
| 1% Phenol Red | 8 ml | 2 ml | 1 ml |
Adjust pH to 7.8-8.2 & Autoclave
| 4 L | 1 L | 0.5 L | |
| Yeast Extract | 400 ml | 100 ml | 50 ml |
| Penicillin (120,000 u/ml) | 20 ml | 5 ml | 2.5 ml |
| Thallium Acetate (1:50) | 100 ml | 25 ml | 12.5 ml |
| Horse Serum | 800 ml | 200 ml | 100 ml |
readjust pH after addition of above!
Mycoplasma Agar
Substitute Agar-PPLO for PPLO broth base
| 4 L | 1 L | 0.5 L |
| 60 g | 15 g | 7.5 g |
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