生物科技行业密歇根大学生物系实验室的常用试剂配方.docx

1、生物科技行业密歇根大学生物系实验室的常用试剂配方（生物科技行业）密歇根大学生物系实验室的常用试剂配方TableofContentsLBMedium.1NZMedium.2SMBuffer.3SETBuffer.46XPrehybSoln.510XTBE.610XTAE.720XSSC.81%SDS,0.2MNaOH.914%PEG(8000),2MNaCl,10mMMgSO4. 1020%SDS.111.0MTris,pH8.0,1.5MNaCl.1210mMTris-HCl,pH7.5,10mMMgSO4.1310mMTris,50mMEDTA,pH7.5.1410mMTris-HCl,1m

2、MEDTA,pH7.5.153MSodiumAcetate,pH4.8.16Electrophoresisdye.17LabellingStopdye.18Sequencinggeldye.195%Acrylamide.206%AcrylamideinTBE,50%Urea.2140%Acrylamide.22LBMedium(1Liter)10gBacto-tryptone5gBacto-yeastextract10gNaClForfortyplatesadd1%agar-1g.Autoclavemedia.Whencool,addampicillinandp ourplates.For1L

3、ofmedia,add1.8mLamp.NZMedium(500mL)5gBacto-tryptone2.5gBacto-yeastextract2.5gNaCl1.25gMgSO4For20platesadd1.2%agar-6g.Autoclaveandpourplatesat50oCSMBuffer(1L)5.8gNaCl1.2gMgSo450mL1MTris-HCl,pH7.50.1gGelatin(doesntdissolve)AutoclaveUsedforphagedilutionandstorage.SETBuffer50mMTris-HCl,pH8.0,50mMEDTA,20

4、%w/vSucrosetomake200mL:40gSucrose10mLof1MTris20mLof0.5MEDTA,disodiumsaltbringto200mLwithH206XPrehybridizationSolutiontomake500mL300mLddH20150mL20XSSC50mL50XDenhardtssolution1mL0.5MEDTA(disodiumsalt)2.5mL20%SDS6XreferstotheconcentrationofSSC10XTBEBuffer(forpolyacrylamidegels)tomakeoneliter:60.75gTris

5、3.7gEDTA(tetrasodiumsalt)30gBoricacid10XTAEBuffer(Foragarosegels)tomakeoneliter:48.20gTris6.75gNaAce3.75gEDTA(disodiumsalt)AdjustpHto7.6withaceticacid.(Approx.20mL)20XSSCtomakeoneliter:175.3gNaCl88.2gNaCitrateaddwatertobringvolumetooneliter.adjusttopH7.0withHCl.1%SDS,0.2MNaOHtomake100mL:93mLddH205mL

6、20%SDS2mL10MNaOH14%PEG(8000),2MNaCl,10mMMgSO4tomakeoneliter:140gPEG117gNaCl2.46gMgSO4ForuseinphageDNApreparation.20%SDStomale250mL:50gofSDSinabeakerAddstirbarandH20last.ThissolutionwillhavetobeheatedfortheSDStodissolve.1.0MTris,pH8.0,1.5MNaCltomakeoneliter:121.1gTrizma87.6gNaClinavolumeofwaterlessth

7、an1L.AdjustpHwithHCl,thenbringto1LwithH2010mMTris-HCl,pH7.5,10mMMgSO4tomakeoneliter:10mL1MTris-HCl2.46gMgSO4foruseinphageDNApreparation10mMTris,50mMEDTA,pH7.5tomake200mL:2mL1MTris20mL0.5MEDTA(tetrasodiumsalt)178mLddH20adjustpHwithHCl.10mMTris-HCl,1mMEDTA,pH7.5tomake200mL:2.0mL1MTris-HCl,pH7.50.4mL0.

8、5MEDTA197.6mLddH203MSodiumAcetate,pH4.8tomakeoneliter:408.1gNaAce(trihydrate;getscoldinsoln)about700mLH20adjustpHwithglacialaceticacid(takesalot)MeasuretrupHbydilutionwithwater;rangewillbebetween4.8and5.5.ElectrophoresisDyetomake4mL:3mL50mMEDTA,10mMTris-HCl,pH8.01mLglycerol20 LBPB10 LXylenecyanolSto

9、pdyeforlabelledprobe1mL50mMEDTA,10mMTris,pH7.5-8.5about200 lglycerol addafewgrainsofbluedextran(8000)Sequencinggeldyeforapprox1mL:1mLformamide10 Lxylenecyanol 10 lBPB3 L10MNaOH5%acrylamidetomake200mL:20mL10XTBE25mL40%acrylamide155mLH206%AcrylamideinTBE,50%Ureatomake500mL:50mL10XTBE75mL40%acrylamide250gUreabringto500mLwithH2O40%Acrylamide(38:2acrylamide:bisacrylamide)tomake200mL:76gacrylamide4gbisacrylamidebringto200mLwithH2O