RatherRapidGenomicPrep
HoffmanandWinston,Gene,1987
1.Grow5mlyeastculturestosaturation.2.CollectcellsbycentrifugationandresUSPendin0.5mlofwater.Transfercellsto1.5mlmicrofugetubeandcollectbya5secondspin.3.Pouroffsupernatantandbrieflyvortextoresuspendcellsinresidualliquid.4.Add0.2mlofBufferA(2%TritonX-100,1%SDS,100mMNaCl,10mMTris-HClpH8.0,1mMEDTApH8.0),200µlglassbeads,and0.2mlphenol:chloroform:isoamylalcohol(25:24:1).5.Vortex3minutes(setting#7onafoammulti-tubevortexadaptor).Add0.2mlTE.6.Spin5minutes.Transferaqueoustonewtube.OPTIONAL:Doachloroformextraction.7.Add1ml100%EtOH(RT;coldEtOHwillcreatealarge,dirtypellet),inverttubetomix,andspin2minutes.8.Discardsupernatant,andresuspendpelletin0.4mlTE(noneedtodrypellet).9.Add10µl4Mammoniumacetate,mix,andthenadd1ml100%EtOHandmix.10.Spinfor2minutesanddrypellet.Resuspendin50µlTEanduse10µlperdigest.