YeastMiniprepfromRobzykandKassir,NucleicAcidRes.(1992)20:3790 Materials: Procedure:•10mlovernightcultureyeastindropoutmedia(mustbeverydense)•STET(8%sucrose,50mMTrispH8,50mMEDTA,5%TritonX-100)•Glassbeads•7.5MNH4OAc•icecoldEtOH•70%EtOH
1)Spindownyeastculture.AspirateoffliquidandresUSPendin100µlSTET.TransfertoscrewtopEppendorftubes.2)Add2scoopsofglassbeads(approx200mg)andvortexfor5min.3)Addanother100µlSTET.Vortexbrieflytoresuspend.4)Boilfor3mininboilingwaterbath.5)Coolbrieflyoniceandspininmicrofugefor10minat4¡.6)Transfer100µlofsupetonewtube.7)Add50µl7.5MNH4OAc.Incubateat-20¡for1hr.8)Spin10minat4¡.(Thisstepgetsridofimpuritiesthatinhibittransformations).9)Collect100µlsupeandaddto200µlicecoldEtOH.10)Spin10minat4¡.11)Rinsepelletwith70%EtOH.12)Resuspendpelletin20µlofwater.Use10µltotransformbacteria.Candoaregulartransformation.
