May 16, 2022
Version 2
Mycology media V.2
- 1KMU
Protocol Citation: Yin-Tse Huang 2022. Mycology media. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvw536wvmk/v2Version created by Yin-Tse Huang
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: May 16, 2022
Last Modified: September 13, 2023
Protocol Integer ID: 62634
Abstract
Mycology media
WA+C agar
Water agar + chloramphenicol (氯黴素 kill bacteria): As an environmental isolating media
| A | B | |
| Component | Amount | |
| Agar | 20g | |
| Chloramphenicol | 0.05g | |
| DI water | 1000 ml |
chloramphenicol is autoclavable
PDA+C agar
potato dextrose agar + chloramphenicol (氯黴素 kill bacteria): Use when WA+C agar is not working
| A | B | |
| Component | Amount | |
| PDA | 39g (or per instruction on label) | |
| Agar | 5g | |
| Chloramphenicol | 0.05g | |
| DI water | 1000 ml |
quarter-strength PDA
1. Less nutrient than regular PDA (1/4)
2. For beetle-associated fungi isolation
| A | B | |
| Component | Amount | |
| PDA | 9.75g (or per instruction on label) | |
| Agar | 5g | |
| DI water | 1000 ml |
MEA (malt extraction agar)
For isolate subculture
| A | B | |
| Component | Amount | |
| MEA | 33.6g (or per instruction on label) | |
| Agar | 5g | |
| DI water | 1000 ml |
Emerson YpSs Agar: For chytrids
| A | B | |
| Component | Amount | |
| Yeast extract | 4g | |
| Soluble Starch | 15g | |
| K2HPO4 | 1g | |
| MgSO4 x 7H2O | 0.5g | |
| Agar | 20g | |
| DI water | 1000ml |
Adujust pH to 7.0 +/- 0.2. Autoclave at 121ºC.
10% Unclarified V8 Agar
- Combine 100 ml V8 juice and 900 ml of distilled water.
- Add 1g CaCO3and 0.05 g b-sitosterol and mix well. Leave stir bar in the flask for later mixing.
- Add 15g of agar.
- Autoclave at 15 psi for 20 minutes.
- Stir medium while dispensing to insure good mixing of CaCO3.
20% : 200 ml V8, 800ml d. water, 2g CaCO3, and 0.05g b-sitosterol.
15% : 150 ml V8, 850 ml d. water, 1.5g CaCO3, and 0.05g b-sitosterol.
Reference: Miller, P. M. 1955. V-8 juice agar as a general purpose medium for fungi and bacteria. Phytopathology 45:461-462.
SceSel+ medium (Kaltseis et al.,2009)
For isolation of Scedosporium
| A | B | |
| Component | Amount | |
| Malt extract | 6.25 g | |
| maltose | 6.25 g | |
| KH2PO4 | 1.25 g | |
| yeast extract | 1 g | |
| Magnesium Sulfate Heptahydrate | 0.625 g | |
| soy peptone | 0.625 g | |
| chloramphenicol | 0.1 g | |
| ciprofloxacin | 0.1 g | |
| streptomycin sulfate | 0.1 g | |
| dichloran | 2 mg | |
| benomyl | 6 mg | |
| DI water | 1000ml |