DNAPlasmidPrep. ThisisascaledupversionoftheBaronprotocolwhichhasbeenmodifiedtoachievepuritycomparabletoCsClpreps.IhavenotsortedoutstrainvariationsyetbutHB101workswell. Solutions SolutionI 1%glucose10gglucose 25mMTrispH8.025ml1MTrispH8.0 10mMEDTA20ml0.5MEDTApH8.0 upto1literwithQ storeat4o SolutionII 1%SDS2.5ml20%SDS 0.2NNaOH1.0ml10NNaOH upto50mlwithQ makefreshasneeded SolutionIII 25%potassiumacetate250gpotassiumacetate add150mlglacialaceticacidandbringupto1liter withQ storeat4o PEG8000 40%PEG8,00040gPEG8,000 upto100mlwithQ storeatroomtemperature Procedure •Spindown5mlofaovernightculturein3Eppendorftubesat14KandpoolthesamplesbyresUSPendingin200mlSolutionIcontaining1mg/mlLysozyme(Sigma#L6876)onice. •Immediatelyadd400mlSolutionII,followedby300mlSolutionIIIandspinat14Kfor5-10". •Followingthespin,remove600-700mlofthesupernatantandadd500mlphenol/chloroform;mixandspinat14Kfor5". •Remove600mlsupernatant,topoffwithcoldEtOHandspin10"atroomtemperature. •Washwith80%EtOH,dryandresuspendin60mlTEplus1mlRNaseA(10mg/ml);incubateat37°for10". •Add50mlphenol/chloroformandspinfor2",remove50mlsupernatantandadd10ml5MNaClplus21ml40%PEG8000. •Incubateonice5",spin5"atroomtemperatureandresuspendin90mlQ(forpUCoriIgetapprox.0.2-0.5mg/ml).