Difference between revisions of "Nourseothricin"
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| − | Nourseothricin is used a dominant resistance marker. We use it mostly in the [[CRISPR Cas9 Full protocol | CRISPR-Cas9]] method for transformation. | + | Nourseothricin is used a dominant resistance marker that is encoded by natMX. We use it mostly in the [[CRISPR Cas9 Full protocol | CRISPR-Cas9]] method for transformation. |
| − | + | ||
| − | Make a stock solution of 100mg/ml | + | Make a stock solution of 100mg/ml |
* dissolving 1 g [https://www.jenabioscience.com Nourseothricin] Nourseothricin-salt in 10ml H<sub>2</sub>O and filter sterilize. | * dissolving 1 g [https://www.jenabioscience.com Nourseothricin] Nourseothricin-salt in 10ml H<sub>2</sub>O and filter sterilize. | ||
* [[filter sterilize]] and aliquot in 1ml tubes. | * [[filter sterilize]] and aliquot in 1ml tubes. | ||
| − | ''' | + | ==Usage== |
| + | '''Pombe''': | ||
| + | |||
| + | Add 1:1000 (i.e. 25µl/plate) to solid medium of ~50-60°C to obtain a final concentration of 100µg/ml. | ||
| + | |||
| + | |||
| + | '''Schizophyllum''' | ||
| + | |||
| + | Add 1:10.000 (i.e. 2.5µl/plate) to solid SMM of ~50-60°C to obtain a final concentration of 10µg/ml. | ||
[[Category:Stock_Solutions]] | [[Category:Stock_Solutions]] | ||
Latest revision as of 16:50, 23 November 2018
Nourseothricin is used a dominant resistance marker that is encoded by natMX. We use it mostly in the CRISPR-Cas9 method for transformation.
Make a stock solution of 100mg/ml
- dissolving 1 g Nourseothricin Nourseothricin-salt in 10ml H2O and filter sterilize.
- filter sterilize and aliquot in 1ml tubes.
Usage
Pombe:
Add 1:1000 (i.e. 25µl/plate) to solid medium of ~50-60°C to obtain a final concentration of 100µg/ml.
Schizophyllum
Add 1:10.000 (i.e. 2.5µl/plate) to solid SMM of ~50-60°C to obtain a final concentration of 10µg/ml.
