McClean: Alpha Factor Stock
Overview
This is the standard stock solution we use for inducing the pheromone pathway in budding yeast. Generally, we find that the different batches of pheromone can be somewhat variable so it is a good idea to make up a stock that will last you through your experiment and keep it in your own -20°C box. If you have to switch stocks mid-experiment you need to do some calibration experiments to make sure the activity is the same.
Materials
α1-Mating Factor acetate salt (Sigma T6901-1mg) DMSO (Fluka 41639, Ultra for molecular biology; stored in the Flammables cabinet)
Procedure
Pipet 1ml of DMSO into the vial of α-factor from Sigma. Pipet up and down and put the lid back on an swirl to dissolve all of the alpha-factor into the DMSO. Then remove the liquid and aliquot it in 100μL eppendorfs into individual eppendorfs and store these in your own -20°C box.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
Contact
- Megan N McClean 14:01, 6 October 2011 (EDT)
or instead, discuss this protocol.
