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Team:Wageningen UR/Experimental protocols
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| - | The standard protocol for media for the SSB group is provided as Appendix D. Media and supplements used here are:<br /> | + | The standard protocol for media for the SSB group is provided as Appendix D. Media and supplements used here are:<br /><br /> |
Uridine solution<br /> | Uridine solution<br /> | ||
| - | Per 100 ml; 12.2 g uridine. Filter-sterilised using a 0.2 µm filter.<br /> | + | Per 100 ml; 12.2 g uridine. Filter-sterilised using a 0.2 µm filter.<br /><br /> |
Vishniac solution<br /> | Vishniac solution<br /> | ||
Per litre; 10 g EDTA; 4.4 g ZnSO4_7H2O; 1.0 g MnCl2_4H2O; 0.32 g CoCl2_6H2O; 0.32 g Cu¬SO4_5H2O; 0.22 g (NH4)6Mo7O24_¬4H2O; 1.47 g CaCl2_2H2O; 1.0 g FeSO4_7H2O. pH adjusted to 4. Filter-sterilised using a 0.2 µm filter.<br /> | Per litre; 10 g EDTA; 4.4 g ZnSO4_7H2O; 1.0 g MnCl2_4H2O; 0.32 g CoCl2_6H2O; 0.32 g Cu¬SO4_5H2O; 0.22 g (NH4)6Mo7O24_¬4H2O; 1.47 g CaCl2_2H2O; 1.0 g FeSO4_7H2O. pH adjusted to 4. Filter-sterilised using a 0.2 µm filter.<br /> | ||
Revision as of 22:11, 11 September 2013
- Safety introduction
- General safety
- Fungi-related safety
- Biosafety Regulation
- Safety Improvement Suggestions
- Safety of the Application
- Lablog
- Experimental protocols
Protocols
Subtitle
Materials
Standard lab procedures are explained in Appendix A, B, C and D. Novel protocols are mentioned and explained at the specific topic pages, since they cannot be explained without the mention of intermediate results and can be considered results. A detailed version of these protocols is also given here.
Strains
A. niger N593
This strain is cspA and pyrA deficient. Media for N593 are supplemented with 5 mM uridine. Unlike citric acid producers, this strain produces mainly oxalic acid.
A. niger strains expressing GFP
Three strains have been applied that express Green Fluorescent Protein (GFP) in various constructs. GFP is widely used as a localisation tool. These strains have been constructed using strain AB4.1 (Vinck, A., M. Terlou, et al. (2005). "Hyphal differentiation in the exploring mycelium of Aspergillus niger." Molecular Microbiology 58(3): 693-699). The GFP constructs are made using the sGFP gene. This mutated GFP variant has an excitation peak at 488 nm and an emission peak at 510 nm. Information is given in table 1.
A. nidulans WG505
This is a pyr-deficient strain, so 5 mM uridine was added to the medium. Use of this strain is mentioned by Nyyssola (Nyyssola, A., R. Heshof, et al. (2012). "Methods for identifying lipoxygenase producing microorganisms on agar plates." AMB Express 2(1): 17). No further data available.
Media
The standard protocol for media for the SSB group is provided as Appendix D. Media and supplements used here are:
Uridine solution
Per 100 ml; 12.2 g uridine. Filter-sterilised using a 0.2 µm filter.
Vishniac solution
Per litre; 10 g EDTA; 4.4 g ZnSO4_7H2O; 1.0 g MnCl2_4H2O; 0.32 g CoCl2_6H2O; 0.32 g Cu¬SO4_5H2O; 0.22 g (NH4)6Mo7O24_¬4H2O; 1.47 g CaCl2_2H2O; 1.0 g FeSO4_7H2O. pH adjusted to 4. Filter-sterilised using a 0.2 µm filter.
