密歇根大学生物系实验室的常用试剂配方

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%acrylam