《密歇根大学生物系实验室的常用试剂配方.docx》由会员分享，可在线阅读，更多相关《密歇根大学生物系实验室的常用试剂配方.docx（10页珍藏版）》请在三一文库上搜索。
1、TableofContentsLBMedium1NZMedium2SMBuffer3SETBuffer46XPrehybSoln510XTBE610XTAE720XSSC81%SDS,0.2MNaOH914%PEG(8000),2MNaCl,10mMMgSO41020%SDS111.0MTris,pH8.0,1.5MNaCl1210mMTris-HCl,pH7.5,10mMMgSO41310mMTris,50mMEDTA,pH7.5143MSodiumAcetate,pH4.816Electrophoresisdye17LabellingStopdye18Sequencinggeldye195
2、%Acrylamide206%AcrylamideinTBE,50%Urea2140%Acrylamide22LBMedium(1Liter)10gBacto-tryptone5gBacto-yeastextract10gNaClForfortyplatesadd1%agar-1g.Autoclavemedia.Whencool,addampicillinandpourplates.For1Lofmedia,add1.8mLamp.NZMedium(500mL)5gBacto-tryptone2.5gBacto-yeastextract2.5gNaCl1.25gMgSO4For20plates
3、add1.2%agar-6g.Autoclaveandpourplatesat50oCSMBuffer(1L)5.8gNaCl1.2gMgSo450mL1MTris-HCl,pH7.50.1gGelatin(doesntdissolve)AutoclaveUsedforphagedilutionandstorage.SETBuffer50mMTris-HCl,pH8.0,50mMEDTA,20%w/vSucrosetomake200mL:40gSucrose10mLof1MTris20mLof0.5MEDTA,disodiumsaltbringto200mLwithH206XPrehybrid
4、izationSolutiontomake500mL300mLddH20150mL20XSSC50mL50XDenhardtssolution1mL0.5MEDTA(disodiumsalt)2.5mL20%SDS6XreferstotheconcentrationofSSC10XTBEBuffer(forpolyacrylamidegels)tomakeoneliter:60.75gTris3.7gEDTA(tetrasodiumsalt)30gBoricacid10XTAEBuffer(Foragarosegels)tomakeoneliter:48.20gTris6.75gNaAce3.
5、75gEDTA(disodiumsalt)AdjustpHto7.6withaceticacid.(Approx.20mL)20XSSCtomakeoneliter:175.3gNaCl88.2gNaCitrateaddwatertobringvolumetooneliter.adjusttopH7.0withHCl.1%SDS,0.2MNaOHtomake100mL:93mLddH205mL20%SDS2mL10MNaOH14%PEG(8000),2MNaCl,10mMMgSO4tomakeoneliter:140gPEG117gNaCl2.46gMgSO4ForuseinphageDNAp
6、reparation.20%SDStomale250mL:50gofSDSinabeakerAddstirbarandH20last.ThissolutionwillhavetobeheatedfortheSDStodissolve.1.0MTris,pH8.0,1.5MNaCltomakeoneliter:121.1gTrizma87.6gNaClinavolumeofwaterlessthan1L.AdjustpHwithHCl,thenbringto1LwithH2010mL1MTris-HCl2.46gMgSO4foruseinphageDNApreparation10mMTris,5
7、0mMEDTA,pH7.5tomake200mL:2mL1MTris20mL0.5MEDTA(tetrasodiumsalt)178mLddH20adjustpHwithHCl.10mMTris-HCl,1mMEDTA,pH7.5tomake200mL:2.0mL1MTris-HCl,pH7.50.4mL0.5MEDTA197.6mLddH203MSodiumAcetate,pH4.8tomakeoneliter:408.1gNaAce(trihydrate;getscoldinsoln)about700mLH20adjustpHwithglacialaceticacid(takesalot)
8、MeasuretrupHbydilutionwithwater;rangewillbebetween4.8and5.5.ElectrophoresisDyetomake4mL:3mL50mMEDTA,10mMTris-HCl,pH8.01mLglycerol20 LBPB10 LXylenecyanolStopdyeforlabelledprobe1mL50mMEDTA,10mMTris,pH7.5-8.5about200 lglyceroladdafewgrainsofbluedextran(8000)SequencinggeldyeforapproxImL:ImLformamide10 Lxylenecyanol10 1BPB3 L10MNaOH5%acrylamidetomake200mL:20mL10XTBE25mL40%acrylamide155mLH206%AcrylamideinTBE,50%Ureatomake500mL:50mL10XTBE75mL40%acrylamide250gUrea bringto500mLwithH2O40%Acrylamide(38:2acrylamide:bisacrylamide)tomake200mL:76gacrylamide4gbisacrylamide bringto200mLwithH2O