May 16, 2023
Version 2
Complete Medium or Complete Medium Xylose (from Leach, Lang and Yoder 1982) V.2
- 1University of Birmingham, School of Biosciences
Protocol Citation: Megan Mcdonald 2023. Complete Medium or Complete Medium Xylose (from Leach, Lang and Yoder 1982). protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg39mxqg25/v2Version created by Megan Mcdonald
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, 2023
Last Modified: May 16, 2023
Protocol Integer ID: 81948
Abstract
For the growth and maintenance of Cochliobolus carbonum and Cochliobolus victoriae
Materials
Trace Minerals
Boric Acid (H3BO3) CAS:10043-35-3
Cupric Sulfate (CuSO4) CAS: 7758-98-7
Potassium Iodide (KI) CAS:7681-11-0
Salts
Potassium Phosphate monobasic KH2PO4 CAS: 7778-77-0
Magnesium Sulfate heptahydrate MgSO4 * 7H2O CAS: 10034-99-8
Sodium Chloride NaCl CAS: 7647-14-5
Media
Yeast Extract CAS: 8013-01-2 (some people report differences between difference sources)
Casein Digests: Still a bit of work to do here to determine, which of these is best (including new plant-based sources). Big differences in prices between different digest types.
Acid hydrolysed Casein CAS: 65072-00-6
Peptone from casein, tryptic digest OR pancreatic digest (tryptone) CAS: 91079-40-2
Make Micronutrients Solution
Make Micronutrients Solution
9 mg H3BO3
58.5 mg CuSO4 *5H2O
1.95 mg KI (Potassium Iodine)
9 mg MnSO4
7.6 mg NaMoO4
822 mg ZnSO4 * 6 H2O
139.8 mg FeCl3 * 6H2O
in 300 mL ddH2O and filter sterilise
Citation:
Heterokaryosis and Parasexuality in the Fungus Ascochyta Imperfecta Author(s): K. E. Sanderson and A. M. Srb Source: American Journal of Botany , Jan., 1965, Vol. 52, No. 1 (Jan., 1965), pp. 72-81 Published by: Wiley Stable URL: https://www.jstor.org/stable/2439977
Make 100x Salt Solutions A and B
Make 100x Salt Solutions A and B
100X Salt Solution A
10g Ca(NaO3)2 * 4 H2O
100 mL ddH2O
Autoclave
Citation:
Leach, J., Lang, B. R. & Yoder, O. C. Microbiology128, 1719-1729, doi:https://doi.org/10.1099/00221287-128-8-1719 (1982).
100X Salt Solution B
2 g KH2PO4 * 7 H2O
1.5g NaCl
100 mL H2O
pH 5.3
Autoclave
Citation:
Leach, J., Lang, B. R. & Yoder, O. C. Microbiology128, 1719-1729, doi:https://doi.org/10.1099/00221287-128-8-1719 (1982).
Make Complete Medium (CM) or Complete Medium Xylose (CMX)
Make Complete Medium (CM) or Complete Medium Xylose (CMX)
Complete Medium Base
10 g glucose OR xylose (substitute glucose for xylose for CMX medium)
1 g Yeast Extract
0.5 g Acid-hydrolysed Casein
0.5 g Enzyme-hydrolysed casein
20g Agar
10 mL Salt A
10 mL Salt B
Make up to 1000 mL with ddH2O and Autoclave
After autoclaving add:
1 mL sterilised micronutrient solution
Citation:
Leach, J., Lang, B. R. & Yoder, O. C. Microbiology128, 1719-1729, doi:https://doi.org/10.1099/00221287-128-8-1719 (1982).
Complete Medium for Sporulation
Complete Medium for Sporulation
Complete Medium Base
0.5 g glucose OR xylose (substitute glucose for xylose for CMX medium)
20 g Sorbose
1 g Yeast Extract
0.5 g acid hydrolysed casein
0.5 g enzyme hydrolysed casein
20g Agar
10 mL Salt A
10 mL Salt B
Make up to 1000 mL with ddH2O and Autoclave
After autoclaving add:
1 mL sterilised micronutrient solution
Protocol references
Citation:
Heterokaryosis and Parasexuality in the Fungus Ascochyta Imperfecta Author(s): K. E. Sanderson and A. M. Srb Source: American Journal of Botany , Jan., 1965, Vol. 52, No. 1 (Jan., 1965), pp. 72-81 Published by: Wiley Stable URL: https://www.jstor.org/stable/2439977
Leach, J., Lang, B. R. & Yoder, O. C. Microbiology128, 1719-1729, doi:https://doi.org/10.1099/00221287-128-8-1719 (1982).