Smolke:Protocols/Dropout plates: Difference between revisions
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==Procedure== | ==Procedure== | ||
N.B. - this protocol makes | N.B. - this protocol makes ~1 sleeve of moderately thick plates. Adjust volumes as needed. | ||
#Mix: | #Mix: | ||
#* | #*400 mL NP water (or less, if using alternate sugars) | ||
#*0. | #*0.85g YNB (Stored in cold room) | ||
#* | #*2.5g NH<sub>4</sub>SO<sub>4</sub> | ||
#* | #*10g Difco agar (''not'' Bacto agar) | ||
#Mix the solution well and autoclave on liquid cycle | #Mix the solution well, add a stir bar, and autoclave on liquid cycle | ||
#After autoclaving, | #After autoclaving, place flask on stir plate to cool | ||
#When bottle is cool enough to touch with the bare hand, add dropout solution and sugar(s) | #When bottle is cool enough to touch with the bare hand, add dropout solution and sugar(s) | ||
# | #Mix, then pour into plates | ||
#*Pour just enough cover the base ( | #*Pour just enough cover the base (without shaking) | ||
#*Briefly flame the top of the plate to eliminate bubbles | #*Briefly flame the top of the plate to eliminate bubbles | ||
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*I've found that it cuts down on condensation if you pile the plates on top of each other while they dry. -[[User:Jkm|Jkm]] | *I've found that it cuts down on condensation if you pile the plates on top of each other while they dry. -[[User:Jkm|Jkm]] | ||
*If you don't want to flame, drop a stirbar into the flask before autoclaving. Mix on the stirplate to add sugar and aa's, stir some more to take care of bubbles. Pour carefully. -[[User:Tsbayer|tsb]] | *If you don't want to flame, drop a stirbar into the flask before autoclaving. Mix on the stirplate to add sugar and aa's, stir some more to take care of bubbles. Pour carefully. -[[User:Tsbayer|tsb]] | ||
*Plate thickness varies greatly depending on who pours the plates. I prefer thicker plates (500mL = 24 plates) rather than thinner plates (500mL = 40+ plates). In my experience, thin plates can dry out before the yeast colonies reach their full growth (so you end up with small colonies and thin streaks). This is particularly problematic for slow-growing strains. | |||
==Contact== | ==Contact== |
Revision as of 14:24, 10 April 2009
OverviewMaking selective yeast plates. Materials
ProcedureN.B. - this protocol makes ~1 sleeve of moderately thick plates. Adjust volumes as needed.
Notes
ContactFor another protocol, look here. |