McClean: Potassium Phosphate

Overview

Potassium phosphate for several procedures including fixing yeast cells for fluorescence measurements.

Materials

• Potassium phosphate monobasic KH₂PO₄
• Potassium phosphate dibasic K₂HPO₄

Procedure

Generally it is easiest to dissolve the entire container of G418 at once (especially if it is a small 1g container).

• Calculate the amount of PBS to add to the vial to get a 200mg/ml 'active' concentration.
  • For example: 710μg/mg means that 710μg out of 1000μg are active, or that the Geneticin is 71% active. Therefore, to make a stock solution of 200mg/ml active Geneticin you need to add 1000/710*200=281.6901mg/ml of Geneticin. The vial contains 1g of Geneticin, so you need to dissolve the contents of the vial in 1000mg/281.6901mg=3.55ml total volume.
• Sterile filter the drug using a syringe and a 0.45μm filter.
• Store stock at -80°C and working stock solution at 4°C

Stock Solutions

Please write in here when you make a new stock solution and how you made it, how many tubes, etc. Also add a note to the "Yeast Media Antibiotic Concentrations" sheet on the front of the 4°C fridge.

*Megan N McClean Made stock on 11/30/2011. Geneticin Lot #919813, activity 701ug/mg. 1000/701*200= 285.3067; 1000/ 285.3067=3.505ml. Dissolve contents of the entire 1g vial in 3.505ml total volume of 1x PBS. Sterile-filted, aliquoted in 500ul in eppendorfs. Left one tube at 4°C and put the others at -80°C

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

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Contact

• Megan N McClean 14:01, 23 September 2011 (EDT)

or instead, discuss this protocol.