1.MakeYPDregularagar(1.5-2%)forsendingstrains. 2.Make200mlbatches=enoughfor6microtiterplates. 3.AddG418at200mg/L. 4.Keepthemediumat65°orhotter.Placethemultichannelpipettorbasinonablockthatissetat65°.Pourthehotmediumnearlytothetop. 5.Dispenseagarmediaintowells.Donotblowoutthefinaldropofmediumintothewells(makesbubbles). ColonyPCR:200µlperwell Priortoinoculatingcells,add10µlwatertoeachwell.Theninoculatethewellswithsteriletoothpicks.Afterallthewellsareinoculated,givetheplateagentleswirltospreadthecellsonthesurfaceofthemediumineachwell.Thismakessemi-liquidculturesforuseastemplateforcolonyPCR,afterwhichtheplatescanbeParafilmedandstored. Sendingstrains:150µlperwell Letplatessitonbenchtodryforseveraldaysbeforeinoculatingwells.DoNOTaddwatertothewells. fromLindaRiles,November1997YPDFILLEDMICROTITERPLATES