[UseddH2O17m*orbetter] 1)LBBroth Make16gmofLBBrothBase(Gibco#M27800C)upto800mlinddH2O. Swirltodissolve,thenadd110µlof10NNaOH.Autoclave. 2)NZYBrothCombine: NaCl4gm(Fisher#S671-500;fromMaterialManag.) MgSO47H2O1.6gm(Sigma#M-2773) YeastExtract4gm(Difco#0127-05-3) N-Z-Amine8gm(Sigma#C-0626) Makeupto800mlinddH2O.Swirltodissolve,thenadd110µlof10NNaOH.Autoclave. 3)TerrificBrothCombine: Tryptone9.6gm(Difco#0123-05-7) YeastExtract19.2gm(Difco#0127-05-3) Glycerol3.2ml(Sigma#G-5516) Makeupto720mlinddH2O.Swirltodissolve,thenautoclave20min. Add80mlof0.17MKPO4(make1.85gmKH2PO4and10gmK2HPO4upto80mlinddH2O;autoclave)andautoclave. 4)LBPlates Make25.5gmofLBAgar(Gibco#M27000C)upto800mlinddH2O. Swirl,thenadd110µlof10NNaOH.Autoclave,thenmixwell. Allowtocoolto50°C(checkwithBioRadtemperatureindicatorstrip)thenadd800µlof50mg/mlampicillinforLB-Amp[red/black],or4mlof10mg/mlkanamycinforLB-Kana[red/red],or1.92mlof5mg/mltetracyclineforLB-Tetra[black/black]. Mixwellbyswirling,thenimmediatelypourontoplatesbyflame.Addjustenoughtocoverbottom.Flamesurfacetoremoveairbubbles. 5)NZYPlatesCombine: NaCl20gm MgSO47H2O8gm YeastExtract20gm N-Z-Amine40gm BactoAgar60gm(Difco#0140-01;fromMaterialManag) Makeupto4000mlinddH2O.Swirl,thenadd900µlof10NNaOH.Autoclavefor75min.Coolto50°C.Pourunderflameonto150mmplates. 6)NZYTopAgarose Add1.33gmagarose(Pharmacia#170554-02)to190mlNZYbroth.Autoclave.Coolto47°C.Solidified,steriletopagarosecanbemicrowavemelted,thencooledto47°C. 7)Antibiotics Ampicillinstock,50mg/mlinddH2O,filtersterilize,aliquotandstoreat-20PreparationofBrothandPlates,etc.