QuickDNAPlasmidPrep Thisprotocolgivesverycleanplasmidprepsforrestrictiondigestsandcloning.However,duetothealkalinelysisstep,theDNAisoftennickedandmaynotgiveexceptionalsequencedata. Solutions TENS 0.1NNaOH1ml10NNaOH 0.2%SDS1ml20%SDS 10mMTris7.51ml1MTris7.5 1mMEDTA200ul0.5MEDTA upto100mlwithQ storeatroomtemperature 3MNaOAc5.2 24.6ganhydroussodiumacetate pHto5.2withaceticacidandbringupto100mlwithQ Procedure •Inoculatea2mlovernightculturewithasinglecolony. •Pellet1.5mloftheovernightcultureandaspirateallbut50mloftheLB. •ResUSPendthecellpelletintheremainingLBandadd300mlTENS. •Vortex,andadd150ml3MNaOAc5.2. •Vortexagainandspinfor2minutesinthemicrofuge.Discardthepelletusingatoothpick. •Add300mlphenol/chloroform,vortexandspinfor5minutesinthemicrofuge. •Retrieve200mlofaqueoussupernatantandadd600mlEtOH.Spinfor10minutesinthemicrofuge,washanddrythepellet. •Resuspendin40mlTE.5mlisgenerallysufficientforrestrictiondigests(addRNaseA).