常用缓冲溶液的配制
来自 : 蚂蚁淘
Alwaystotrytopreparebuffersolutionatthetemperatureandconcentrationbeforeplaningtouseduringtheexperiment.StocksolutionsareacceptableaslongaspHadjustmentmadeaftertemperatureandconcentrationadjustment.Alsogoodbuffers,e.g.,TrisandPhosphate,arestableforalongtimeperiod.1X1L10X10mMTris-HCl8.9gTris-HCl5.3gTris-Base2mMNa3EDTA7.4gpH8at25C.Mixandstoreat4C.Periodicallydumpthe1XTEandmakeupthefresh1XTEfrom10Xstock.1X1L1X1L10X10mMTris-HCl1.06gTris-HCl10.6g0.39gTris-Base3.94g1mMNa2EDTA0.475g4.75gpH8at25C.Mixandstoreat4C.1X1L10X50mMTris-HCl44.4Tris-HCl26.5Tris-Base50mMNa3EDTA186gStoreat4C.1X0.25L1X25mMTris-HCl0.55gTris-HCl0.33gTris-Base10mMNa3EDTA0.92g50mMGlucose2.25gStoreat4C.50mM1MTrispH8.05ml1mM0.5MEDTA2mlH2Oupto100mlStoreat4C.1X1L1X2L1X1MTris-HCl88.8gTris-HCl178g53.0gTris-Base106g1.5MNaCl87.7g175gStoreat25C(roomtemperatureisOK)1X1L1X2L1X1MTris-HCl132.2gTris-HCl264.4g19.4gTris-Base38.8g1.5MNaCl87.7g175gStoreat25C(roomtemperatureisOK)Conc.1L2L3L0.5MTrisTris-HCl63.5g127.0190.0Tris-Base11.8g23.6g35.4g1.5MNaCl87.6g175.0g262.8gStoreat25C(roomtemperatureisOK)Conc.StockVolume10mM1MTrispH7.5100µl15mM5MNaCl30µlH2Oto10mlStoreat4C.1X1L10X4L10X89mMTris-Base108g432g89mMBoricacid55g220g2mMNa2EDTA9.3g37.2gEtBr(10mg/ml)0.5ml2mlMixandstoreatroomtemperaturewithoutEtBr,4CwithEtBrinabrownbottle.UseEtBrstocksolution(10mgEtBr/ml)whenTBEismade.1X1L50X1L10X40mMTris-acetate242gTrisBase48.4g57.1mlaceticacid11.4ml2mMNa2EDTA37g7.4gEtBr(10mg/ml)2.5ml0.5mlMixandstoreatroomtemperaturewithoutEtBr,4CwithEtBrinabrownbottle.UseEtBrstocksolution(10mgEtBr/ml)whenTAEismade.Conc.Stock1L1X100mMNa2citrate.2H2O29.4gGlucose63g5mM0.5MNa2EDTA10mlNa2Diethyldithiocarbamicacid10gPVP-40,000MW20gBSA10gAdjusttopH6.0withHCl.Storeat4C.Conc.StockVolume(ml)10mM1MTrispH8.01.02mM0.5MEDTA0.40.2%20%SDA1.0H2O97.6Storeatroomtemperature.Conc.Stock1X1L10X1L10mM1MTrispH8.010.08.88gTris-HCl5.3gTris-Base1mM0.5MEDTA2.04.65gNa2EDTA100mM5MNaCl20.05.84gNaClH2Oupto1Lupto1LpH8.0at25C.Storeat4C.1X1L1X2L1X0.5NNaOH20.0g40g1.5NNaCl87.7g175gStoreatroomtemperature.Conc.1L2L3L4L0.2NNaOH8.0g16.0g24.0g32.0g0.6MNaCl35.04g70.08g105.2g140.2gStoreatroomtemperature.1X1L20X2L4L6L150mMNaCl175.3g350.6g701g1052g15mMNa3Citrate88.3g176.6g353g530gMix,adjustpHto7.0withHCl,andstoreat4C25X1L4L3.0MNaCl219g876g0.3MNa2Citrate110g440gMix,adjustpHto7.4withHCl,andstoreat4C.1X200ml1X3MKAc29.4g(or20mlof5MKAc)2MAceticacid23mlDissolvetheKAcin150mlofH2O,bringto177ml,andthenaddtheglacialaceticacid.Mixandstoreat4C.Conc.Stock1L2L10%50%PEG2004007%20%SDS3507000.6X25XSSC244810mM1MNaHPO410205mM0.5MEDTA1020100µg/mlDenatureSalmonSperm1020H2O396792Aliquote45mlinto50mlPPT,storeat-20C.Afterpreparingthefollowingstocksolutions,mixA,B,andCinaratioof1:2.5:1.5,repectively.Stock2-mercaptoethanol(bME)18µlDXTPs(A,T,G)5µleachSolutionO850µlStoreat-20CConc.Stock250ml100ml1.47MTris-Base44.52g17.81g0.147MMgCl27.47g2.99gAdjustpHto8.0withconc.HCl.:2MhepesbufferConc.Stock250ml100ml2MHepes119.15g47.66gAdjustpHto6.6with4NNaOH.Storeat4C.Hexameroroligonucleotides.Add55µlH2OdirectlytothePharmaciabottlewhichcontains50unitsoflyophilizedhexamer.Storeat-20C.Diluteto1unitKlewnowfragmentbyaddingklewnowbuffer.StockKlenowfromBRLcomesas500unitsin84µl.Diluteto1unitbyadding420µlofklenowbuffertothe500unit/84µlstock.Store-20C.ConcStock7mMTris-HCl0.211g7mMMgCl20.355g50mMNaCl0.730g50%Glycerol125mlAddtheabovetoabout80mlH2O.Stirtodissolve.Adjustvolumeto250mlwithH2O.Storeat-20C.1XConc.Stock500mM1MTris(pH8.0)2.5ml100mM1MMgCl20.5ml500mMNaCl0.5ml25mMSpermidine(3HCl)0.032gH2O1.5mlMix,aliquote2mlinscrew-capvial,storeat-20C.1XConcStock500mM1MTris(pH8.0)2.5ml100mM1MMgCl20.5ml1M5MNaCl1.0ml25mMSpermidine(3HCl)0.032gH2O1.0mlMix,aliquote2mlinscrew-capvial,storeat-20C.
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