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Appendix G
Media Glossary
January 2008
Glossary of agars, broths and other reagents
Section S
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(PDF Version - 35 K)| Basal Medium | |
|---|---|
| Dextrose | 40.0 g |
| Peptone | 10.0 g |
| Agar | 15.0 g |
| Distilled water | 1.0 L |
| pH 5.6 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Autoclave at 121oC for 15 min. Do not over autoclave. Dispense into sterile Petri dishes.
| Working Solution | |
|---|---|
| Safranin | 0.5 g |
| Distilled water | 100 mL |
Complete Medium:
Dissolve 0.5 g of Safranin in 100 mL distilled water.
| Stock Solution | |
|---|---|
| Safranin O (certified) | 2.5 g |
| Ethanol (95%) | 100 mL |
| Working Solution | |
| Stock solution | 10.0 mL |
| Distilled water | 90.0 mL |
| Working Solution | |
|---|---|
| Sodium chloride (NaCl) | 8.5 g |
| Distilled water | 1.0 L |
Complete Medium:
Suspend sodium chloride in 1.0 L of distilled water. Stir to dissolve. Dispense into appropriate glassware and autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Tryptone | 5.0 g |
| Lactose | 4.0 g |
| Sodium phosphate dibasic (Na2HPO4) | 10.0 g |
| Sodium selenite (Na2SeO3) | 4.0 g |
| L-Cystine | 0.01 g |
| Distilled water | 1.0 L |
| pH 7.0 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat for 10 min in autoclave on flowing steam or boil on a hotplate with frequent swirling. DO NOT AUTOCLAVE. Cool to 45-50oC. Dispense into appropriate glassware. Use medium on the day of preparation.
Note: Selenium salts can be teratogenic and should be weighed in a safety cabinet.
| Basal Medium | |
|---|---|
| Newborn calf serum (sterile) | 9 Parts |
| Myo-inositol (50% wt./vol.) | 1 Part |
Complete Medium:
Mix ingredients together well.
| Basal Medium | |
|---|---|
| Tryptone | 20.0 g |
| Potassium phosphate dibasic (K2HPO4) | 2.0 g |
| Potassium phosphate monobasic (KH2PO4) | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Glucose | 1.0 g |
| Tween 80 | 1.5 mL |
| Distilled water | 1.0 L |
| pH 7.0 ± 0.2 | |
| Supplement | |
| Novobiocin | 50 mg |
| Distilled water | 1.0 L |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Autoclave basal medium at 121oC for 15 min. Cool to 50oC. Aseptically add 2.5 mL of novobiocin solution to 225 mL of basal medium and mix thoroughly.
Novobiocin solution : Suspend 50 mg of novobiocin into 1 liter of distilled water. Sterilize by filtration through 0.45 : m membrane filter.
| Basal Medium | |
|---|---|
| Peptone | 30.0 g |
| Beef extract | 3.0 g |
| Iron peptone | 0.2 g |
| Sodium thiosulfate (Na2S2O3) | 0.025 g |
| Agar | 3.0 g |
| Distilled water | 1.0 L |
| pH 7.3 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Dispense 6.5 mL into screw-cap tubes and autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Sodium chloride ( NaCl ) | 5.0 g |
| Magnesium sulfate (MgSO4.7H2O) | 0.2 g |
| Ammonium phosphate (NH4H2PO4) | 1.0 g |
| Potassium phosphate dibasic (K2HPO4) | 1.0 g |
| Sodium citrate (Na3C6H507.2H20) | 2.0 g |
| Bromthymol blue | 0.08 g |
| Agar | 15.0 g |
| Distilled water | 1.0 L |
| pH 6.9 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Dispense 10 mL into culture tubes. Autoclave at 121oC for 15 min. Slope to give 1 inch butts and let cool.
| Basal Medium | |
|---|---|
| Instant nonfat dry milk | 100 g |
| Distilled water | 1.0 L |
| Brilliant green dye solution, aqueous (0.1%) | 20 mL |
| Supplement | |
| Brilliant green dye | 0.1 g |
| Distilled water | 100 mL |
Complete Medium:
Suspend instant nonfat dry milk in 1.0 L of distilled water and stir to dissolve. Add 20.0 mL of 0.1% (w/v) aqueous brilliant green solution (final concentration of 0.002% w/v). Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Distilled water | 1.0 L |
| pH 7.6 | |
Complete Medium:
Suspend peptone in 1.0 L of distilled water and stir to dissolve. Adjust pH to 7.6. Dispense 4.7 mL into culture tubes. Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Sodium chloride ( NaCl )* 3% | 30.0 g |
| 7% | 70.0 g |
| 10% | 100.0 g |
| Distilled water | 1.0 L |
| pH 7.6 | |
Complete Medium:
Suspend peptone in 1.0 L of distilled water and stir to dissolve. Adjust pH to 7.6. Dispense 4.7 mL into culture tubes. Autoclave at 121oC for 15
*For 7% and 10%, use 70 g and 100 g of NaCl, respectively.
| Basal Medium | |
|---|---|
| Trisodium citrate (Na3C6H5O7.2H2O) | 20.0 g |
| Distilled water | 1.0 L |
| pH 7.5 ± 0.2 | |
Complete Medium:
Dissolve 20.0 g trisodium citrate dihydrate in 1.0 L of distilled water by heating to 45-50°C. Adjust to the desired pH. Dispense 99.0 (90.0) mL amounts into dilution blanks. Autoclave at 121°C for 15 min.
| Basal Medium | |
|---|---|
| Sodium phosphate dibasic anhydrous (Na2HPO4) | 8.23 g |
| Sodium phosphate, monobasic, monohydrate ( NaH2PO4 - H2O ) | 1.2 g |
| Sodium chloride (NaCl) | 5.0 g |
| Bile salts ( No. 3 Difco ) | 1.5 g |
| D-Sorbitol | 10.0 g |
| Distilled water | 1.0 L |
| pH | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Peptone | 17.0 g |
| Proteose peptone | 3.0 g |
| Sorbitol | 10.0 g |
| Bile salts No. 3 | 1.5 g |
| Sodium chloride (NaCl) | 5.0 g |
| Neutral red | 0.03 g |
| Crystal violet | 0.001 g |
| Agar | 15.0 g |
| Distilled water | 1.0 L |
| pH 7.1 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Autoclave at 121oC for 15 min. Cool to 45-50oC. Dispense into sterile Petri plates.
| Basal Medium | |
|---|---|
| Sorbitol MacConkey Agar (see above) | 1.0 L |
| (BCIG) 5-bromo-4-chloro-3-indolyl- β -D-glucuronide-Na (or -CHX) salt | 0.1 g |
Complete Medium:
Prepare Sorbitol MacConkey agar and add BCIG-Na salt. Autoclave at 121oC for 15 min. Cool to 50oC . Dispense into sterile Petri dishes.
Note: If using BCIG-CHX, it must be in solution, as follows: dissolve 0.1 g BCIG in 2.5 mL 95% ethanol and 0.5 mL of 1N NaOH. Heat slightly to dissolve and add to 1.0 L Sorbitol MacConkey agar.
| Basal Medium | |
|---|---|
| Sudan black B | 0.3 g |
| Ethanol, 70% | 100 mL |
Complete Medium:
Dissolve Sudan Black B in 70% ethanol.
| Basal Medium | |
|---|---|
| Tryptose | 15.0 g |
| Soytone | 5.0 g |
| Yeast extract | 5.0 g |
| Ferric ammonium citrate | 1.0 g |
| Sodium metabisulfite (Na2S2O5) | 1.0 g |
| Agar | 20.0 g |
| Distilled water | 1.0 L |
| pH 7.6 ± 0.2 | |
| Supplement | |
| D-cycloserine (use white crystalline powder only) | 0.4 g |
| Distilled water | 10 mL |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling to dissolve completely. Autoclave at 121oC for 15 min and cool to 45-50oC. Aseptically add 10.0 mL of the 4% D-cycloserine solution and mix thoroughly. Dispense into sterile Petri dishes.
4% D-cycloserine solution : Suspend 0.4 g of D-cycloserine into 10 mL of distilled water. Sterilize by filtration through 0.45 : m membrane filter.
Note: Agar base is commercially available from BD BBL (SFP Agar Base Shihidi Ferguson Perfringens and TSC Agar Base) and Oxoid (Perfringens Agar Base).
