Smolke:Protocols/Dropout plates

Overview
Making selective yeast plates.

Materials

 * Nanopure water
 * Yeast nitrogen base (YNB)
 * Ammonium sulfate
 * Agar
 * Sugars (normally 20% dextrose, alternative 20% raffinose/10% sucrose/20% galactose)
 * Amino acid dropout solution (in 4&deg;C room)

Procedure
N.B. - this protocol makes ~1 sleeve of moderately thick plates. Adjust volumes as needed.


 * Mix:
 * 1) *400 mL NP water (or less, if using alternate sugars)
 * 2) *0.85g YNB (Stored in cold room)
 * 3) *2.5g NH4SO4
 * 4) *10g Difco agar (not Bacto agar)
 * 5) Mix the solution well, add a stir bar, and autoclave on liquid cycle
 * 6) After autoclaving, place flask on stir plate to cool
 * 7) When bottle is cool enough to touch with the bare hand, add dropout solution and sugar(s)
 * Mix, then pour into plates
 * 1) *Pour just enough cover the base (without shaking)
 * 2) *Briefly flame the top of the plate to eliminate bubbles

Contact
Win

For another protocol, look here.