COMPLEXMEDIA YPADmediumorAlsoCalledYPDplusAdenineYeastExtract-Peptone-DextroseplusAdeninemedium 6.0gmyeastextract(Difco)12.0gmPeptone(Difco)12.0gmGlucose60mgadeninehemisulphate600mldistilledwaterBacto-agar(Difco)10g(addedforagarmedium) Sterilisebyautoclavingat121°Cfor15min. Wealwaysusea1literflaskandmake600mlsofmediabecauseitgivesexactly20plates(1Sleeveofplates).Inadditionthisvolumeofliquidismoreeasilyhandledwhenpouringafterautoclaving. SYNTHETICCOMPLETEDROPOUT(SCdrop-out)MEDIUM 4.0gmDifcoYeastNitrogenBase(w/oaminoacids)4.0gm12.0gmglucose0.50gmSyntheticCompleteDropOutMix600mldistilledwater10.0gmDifcoBactoAgar(addforsolidmedium) Addtheingredients(exceptAgar)tothewaterandmixandadjustthepHto5.6with10NNaOH.(THISSTEPISIMPORTANTFORhighestefficiencyTRANSFORMATION)Autoclaveat121°Cfor15min. SyntheticCompletedrop-outMediumMix(SCdrop-out) SupplementAmountNotethehintaboutmakingthismediabelow 2.0gmAdeninehemisulfate*2.0gmArginineHCl2.0gmHistidineHCl*2.0gmIsoleucine2.0gmLeucine*2.0gmLysineHCl2.0gmMethionine3.0gmPhenylalanine6.0gmHomoserine3.0gmTryptophan*2.0gmTyrosine1.2gmUracil*9.0gmValine Omittheappropriatecomponent(s),indicatedinboldtype,toprepareSC-Ade,SC-His,SC-Leu,SC-Trp,SC-Trp/Leu,andSC-Trp/Leu/His.Combinetheingredientsinaclean250mlplasticbottle.Addthreeorfourcleanglassmarblesandshakevigorouslytomix.Thisquantitywillbesufficientforapproximately100(600ml)batchesofSCdrop-outmedium.Somerecipescallfor20aminoacidsintheSCmixturehowevermostyeaststrainsdonotneedallofthem.Inadditionsomerecipesdonotusehomoserineduetotheexpense,itcanbereplacedbytheadditionofserineandthreonine(2gramseach).
