| |||||||
FastFred's Media CookbookbyFastFred & ChesterKarma(v0.97) polish transl. http://www.psilosophy.info/oudfudjpatezadbxcpavcien
original source: http://www.shroomery.org/forums/showflat.php/Number/3077006 Table of Contents: Section A. Classic Favorite RecipesPotato Dextrose Agar (PDA)(FDA M127)200 g Potato infusion [dilute to 1L total, ~4g solids] To prepare potato infusion, boil 200 g scrubbed, sliced(unpeeled) potatoes in 1 liter distilled water for 30 min. Filter through cheesecloth, saving effluent, which is potato infusion (or use commercial dehydrated form). Mix in other ingredients and boil to dissolve. [Dilute to obtain 1 L final volume] Autoclave 15 min at 121°C. Dispense 20-25 ml portions into sterile 15 x 100 mm Petri dishes. Final pH, 5.6 ± 0.2. Medium should not be re-melted more than once. Medium powder is available commercially but may require supplementing with extra agar to a final concentration of 20 g/liter. To BBL or Difco dehydrated medium, add 5 g of agar. The broth is clear to slightly opalescent and yellowish in color.1, 2 Potato Dextrose Yeast (PDY)200 g Potato, infusion from [dilute to 1L total] Gently boil for ten minutes or until the solution is clear. Malt Extract Agar (3%)(MEA)(General Microbiology)(FDA M93)30 g Malt extract Boil to dissolve ingredients. Avoid overheating, which causes softening of agar and darkening of medium color. Autoclave 15 min at 121°C. Dispense 20-25 ml into sterile 15 x 100 mm Petri dishes. Final pH, 5.5 ± 0.2. This medium is recommended as a general maintenance medium.1 Malt Extract Agar with Yeast (2%)(MEAY)20 g extra light malt extract Sabouraud's Dextrose Broth and Agar (SabDex)(FDA M133)40 g Dextrose Dissolve completely and dispense 40 ml portions into screw-cap bottles. Final pH, 5.8. Autoclave 15 min at 118-121°C. Do not exceed 121°C. For Sabouraud's dextrose agar, prepare broth as above and add 15-20 g agar, depending on gel strength desired. Final pH, 5.6 ± 0.2. Dispense into tubes for slants and bottles or flasks for pouring plates. Autoclave 15 min at 118-121°C.1 Sabouraud Dextrose (SabDex) Agar is used for the isolation, cultivation, and maintenance of saprophytic and pathogenic yeasts and fungi. SabDex Agar is an excellent substitute for Malt or Potato Dextrose Agar, when used by mushroom cultivators to propagate mushroom mycelium. Sabouraud Dextrose Agar was described by Sabouraud in 1892 and was used for the identification of fungi based on their morphological characteristics. Sabouraud Dextrose Agar is a standard medium used to support the growth of yeasts and molds. It supplies peptone as the protein source and dextrose as the carbohydrate source for nourishment. Bacterial suppression occurs due to the low pH. This media is especially suited for the primary isolation of fungi from normally sterile sites such as cerebrospinal fluid (CSF). Later, Emmons modified the medium by decreasing the dextrose content and adjusting the pH closer to the neutral range. This modification enhances sporulation and is particularly useful for the subculture of fungi that so not develop fruiting structures on other media, and so is useful in their identification. It also serves as a good holding medium for stock cultures.3 Section B. Special Blends"Karo Water" (Corn Syrup Broth)1 Teaspoon Light Corn Syrup (Karo Syrup or store brand Light Corn Syrup) Mix well until dissolved, sterilize for 20-30 minutes at 15 psi.17 "Dextrose Tek" Liquid Media (Corn Sugar Broth)1 Teaspoon Powdered Dextrose (corn sugar) Honey Water (Mycotopia Honey Tek)40 g Honey (or roughly 1 tablespoon per pint of water) The correct mixture for optimum results is 4% sugars (honey) by weight and 96% water. Malt-Yeast-Peptone Agar (McKenna's MYP)7 g malt extract (powdered or syrup) Malt Yeast Peptone Agar (Stamets MYP)20 g extra light malt extract Grey Cardboard (instead of agar)Here are the detailed steps for making cardboard plates. Note that you can also use small jars in place of Petri dishes. 1) Measure about 100 mls. of tap water into a small jar. Oatmeal Flake Agar75 g Oatmeal flakes Stir for 5-10 minutes then filter out the larger particles by pouring it through some mesh, save the broth. [Then add the agar] This is the best medium for Panaeolus cyanescens I've ever encountered. Beware, this medium will be less firm than the other recipes so extra agar has to be added to compensate.22 Moonflower's Rice Malt-Alfalfa-Brewer's Yeast Agar1 cup alfalfa, infusion from Prepare infusion using approx. 1 1/2 quarts of clean water. Mix in the alfalfa and rice, then allow to soak for 2 hours at room temp with occasional stirring. Filter or strain before adding the infusion. MycoPsycho's Liquid Mycelial Culture Broth1/2 tsp malt (2.5 ml) 1. procure a half-pint jar with lid & ring and also a spore syringe. Ragadinks Liquid Medium16.5 g dextrose Amaranth Soy Agar20 g amaranth flour EntheoGenesis No.44210 g amaranth flour Sterilize for 20-30 minutes at 15 psi.20 Section C: Food Based Recipes and VariationsCorn Meal Agar (CMA)2 g Corn Meal, Infusion from [filter] Cornmeal Dextrose Agar25 g yellow cornmeal Potato Flake Agar20.0 g Potato Flakes Potato Dextrose Agar and Potato Flake Agar are formulations developed to promote sporulation of fungi. The potatoes contained in these media provide nutritious bases for luxuriant growth of fungi. Both media contain dextrose as a growth stimulant. Potato Starch Agar30 g potato starch, soluble The pH is adjusted following autoclaving to prevent agar hydrolysis by acid.20 Potato Flake Agar with Yeast20 g potato flakes Potato-Carrot Agargrated potato 20.0 g Boil potato and carrot in 1000.0 ml of water for 1 h, filter, add water to the initial volume, adjust pH to 7.0 - 7.1 and add agar. Barley Flour Malt Extract Agar40 g barley flour Sterilize for 20-30 minutes at 15 psi.21 Barley Flour Modified Sabouraud's25 g barley flour Sterilize for 20-30 minutes at 15 psi.21 Oatmeal Agar (OA)60.0 g Oatmeal [filter] Cook oatmeal 5-10 minutes then filter the liquid into another container using cheesecloth or a metal strainer with a tight mesh. Dilute liquid to 1L, add agar, and heat with swirling until solids dissolve. This is reported to be a good media for cultivating Panaeolus cyanescens.5 Oatmeal Agar AOatmeal 20.0 g pH 7.2.6 Oatmeal Agar BOats 30.0 g Keep oats on a water bath at 58°C for 1 h, filter through 2 layers of gauze, dilute V-8 Oatmeal Agar50 ml V-8 juice Be careful to use a container much larger then the volume of medium, i.e., prepare a 500 ml medium in 2 liter flasks or it will tend to boil over no matter how slowly it is cooled down.20 V8 Medium50 ml V8 Juice The commercial V8 juice is occasionally used for tissue cultures of edible mushrooms. It should be noted that most mushrooms prefer neutral to slightly acid range of medium, that is, a pH of about 5.5 to 6.5. However the straw mushroom, Volvariella volvacea, prefers a high pH medium, 6.8 to 7.8. Therefore, it is important to make sure the acidity or pH of the medium is correct for a particular mushroom. Here be careful to use a container much larger then the volume of medium, i.e., prepare a 500 ml medium in 2 liter flasks or it will tend to boil over no matter how slowly it is cooled down. 20 Bean AgarBeans (peas or pulse) 100.0 g [infusion from (filter)] Prepare infusion from beans. Sterilize at 121°C for 30 min.6 Faba Bean Dextrose Agar (FDA)200 g faba bean seeds or 400 g of faba bean leaves [infusion from] Method: This medium is used for propagation of B. fabae, A. fabae, and A. tenuis.12 Pea Agar100 g Yellow peas Cabbage AgarCabbage 50.0 g Boil 50.0 g of cabbage in 1000.0 ml of water, filter cabbage, adjust the volume of broth to the initial value.6 Dr. Pollock's Modified [Dog Food] Agar10 g dried dog food (ground to flour) Sterilize for 20-30 minutes at 15 psi.20 Section D. Other Media and Alternate FormulationsPotato-Glucose Agar 1grated potato 200.0 g Boil potatoes for 1 h in 1 L of water, filter through gauze. Add water to the initial volume, add glucose and agar. Potato-Peptone MediumPotato decoction 200 ml [infusion from 200 g potato] Potato-Peptone-Yeast Agar (PPYA)Potato decoction 200 ml [infusion from 200 g potato] pH 8.0.6 Malt Agar (MA)(FDA M185)[aka 2% MEA]20 g Malt extract, powdered Mix ingredients, steam to dissolve agar and sterilize for 15 min at 121°C. Temper medium to 45°C and pour plates under aseptic conditions. This medium is recommended as a general maintenance medium.1 Difco Malt Extract Broth (FDA M94)6.0 g Malt extract base Final pH, 4.7 ± 0.2.1 Malt Extract Agar for Yeasts and Molds (MEAYM)(FDA M182)20.0 g Malt extract, powdered Mix ingredients, heat to dissolve agar and sterilize at 121°C for 15 min. Temper media to 45°C and pour plates under aseptic conditions. Dehydrated MA is commercially available, but since several MA formulas exist, check for the correct composition. Malt Extract Peptone Agar30 g Malt extract Adjust pH to 5.6. Sterilize at 121°C for 10 min.8 Raper & Thom MEA (RTMEA)To 2% MEA add: ISP 2 Medium (Malt, Yeast, Glucose)Malt extract 10.0 g pH 7.2.6 Yeast Extract Agar (YEA)(FDA M181)10.0 g Proteose peptone Adjust pH to 7.2-7.4. Autoclave at 121°C for 15 min.1 Yeast Glucose AgarYeast extract 5.0 g pH 7.2. Sterilize at 121°C for 15 min.6 Glucose and Yeast Extract AgarGlucose 20.0 g Glycerin Yeast AgarYeast extract 5.0 g Manure TinctureCow manure (fresh) 1.0 kg Boil, squeeze through gauze into a bottle and dilute 3 to 1.6 Manure AgarHorse manure 100-125 g Boil manure in 1000.0 ml of water for 10 min, then keep for 16-20 h, filter through 1-2 layers of filter paper, adjust to the initial volume, add agar. Water Agar (aka Starved Agar)Agar 20.0 g Sterilize at 121°C for 15 min.6 Gelatin Agar (GA)(FDA M54)4 g Peptone Suspend ingredients with constant stirring to prevent scorching gelatin, and boil to dissolve gelatin and agar. Adjust to pH 7.2 ± 0.2. Autoclave 15 min at 121°C. Cool to 45-50°C. Pour plates.1 Plate Count Agar (SMA)(aka Standard Methods Agar)(FDA M124)5.0 g Tryptone Heat to dissolve ingredients. Dispense into suitable tubes or flasks. Autoclave 15 min at 121°C. Final pH, 7.0 ± 0.2. For viable yeasts and molds, dispense 20-25 ml portions into sterile 15 x 100 mm Petri dishes.1 Nutrient Agar (FDA M112)3 g Beef extract Heat to boiling to dissolve ingredients. Dispense into tubes or flasks. Autoclave 15 min at 121°C. Final pH, 6.8 ± 0.2.1 Starch Agar (FDA M143)(Nutrient agar with starch)23 g Nutrient agar (FDA M112) Heat to dissolve agar in 500 ml water. Dissolve starch in 250 ml water. Combine and dilute to 1 liter. Autoclave 15 min at 121°C. Note: add 3 g agar to Difco's dehydrated starch agar.1 Starch-Yeast AgarYeast extract 2.0 g pH 7.3.6 1/5 Starch-Yeast AgarYeast extract 0.4 g pH 7.3.6 Long-term Preservation Medium (FDA M85)3 g Yeast extract, 0.3% Heat to dissolve ingredients. Dispense 4 ml portions to 13 x 100 mm screw-cap tubes. Autoclave 15 min at 121°C. Cool and tighten caps for storage. No pH adjustment is necessary.1 Peptone Meat Agar (Meat Water)Peptone 10.0 g Preparation of meat water: comminute 500 g of meat free of bones, fat and tendons, add 1000.0 ml of tap water and leave for 12 h at room temperature or in a thermostat at 30°C, or for 2 h at 37°C. Then squeeze the meat through gauze or cloth and boil the filtrate for 5 min. The proteins are denatured. Filter the cooled down mass through a cotton-wool filter and add water to the initial volume. pH 7.2 - 7.4. Section E. Common SolutionsGentamicin Sulfate Solution (FDA M57)5.00 g Gentamicin sulfate Sterilize by filtration through 0.20 µm membrane. Store at -20°C.1 Chloramphenicol 0.2g/liter of media survives autoclaving. Section F. Sources
| |||||||