PDF-Proc.Nat.Acad.Sci.USAVol.73,No.4,pp.1029-10&3,April1976BiochemistryTra

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1030BiochemistrySchultzandHallSolutionsExtractionbufferwas02MTrisClpH7922020volvolglycerol20mMMgCl208MNH42SO410mMEDTAand10mMdithiothreitolImmediatelybeforeusephenylmethylsulfo

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Proc.Nat.Acad.Sci.USAVol.73,No.4,pp.1029-10&3,April1976BiochemistryTra: Transcript

1030BiochemistrySchultzandHallSolutionsExtractionbufferwas02MTrisClpH7922020volvolglycerol20mMMgCl208MNH42SO410mMEDTAand10mMdithiothreitolImmediatelybeforeusephenylmethylsulfo. Proc.Natl.Acad.Sci.USA87(1990)wereprepared(25),andtheDNAinsertwasexcisedwithEcoRI,isolatedbylowmeltagarosegelelectrophoresis,andrandom-primedforexpressionanalysis.Theexpressionlev-elsandsizesofthemRNA Proc.Natl.Acad.Sci.USA73(1976)29290.6OD0.50.41+mPMU0.3LDEPTH(mm)IIIIIIII00.51.0FIG.1.Atracedcomputeroutputdisplayingtheaveragedopticaldensitiesoflayersinthespecimenofcerebralcortexshownbelow.Thescanni Proc.Nat.Acad.Sci.USA91(1994)9761thepGEM4vector(Promega)forsequencing.Specifically,a1.2-kbPstIfragment,whichhybridizedtothemousecDNAprobe,butnottotalmouseDNA,wassubclonedtogeneratethesequencehomologou Proc.Natl.Acad.Sci.USA75(1978)RESULTSWhennormalhumanglobinDNAwasdigestedwithEcoRIandprobedwithamixtureofa-and(3-globincDNAs,fivemajorfragmentswereseen(Fig.1).Thelargestfragment,about21kilobases(kb)inl Proc.Natl.Acad.Sci.USA82(1985)855timeswiththesamebufferlackingNP-40andTritonX-100andwereresuspendedinthenick-translationbufferasde-scribedabovecontaining100,Ciof[3H]TTP(117Ci/mmol,NewEnglandNuclear)in Proc.Nat.Acad.Sci.USA73(1976)951Themutagenicpotencyofeachchemicalisbasedonthere-versionofthattesterstrainmostsensitivetoit;eachstrainhasbeenindependentlyoptimizedfordetectionofapartic-ularclassofmutag Proc.Nat.Acad.Sci.USA70(1978)624~~~~~~~~~~~~24*0aS5!!402040608000020-tO60eo0as--40506~~~~~~~~~~~~~0GROWTHINTHEL00H000)CRW'0NTE0~Z04-20FIG.2.(A)Kineticsofinductionofthechloroplastisoleucyl-tRNAuponexpo Proc.Natl.Acad.Sci.USA93(1996)11529BothLandL*canbeprotonatedonLys-156,butthepKaforprotonationisconsiderablylowerforL*thanforL;pKL*iswhilepKLis10.OneimportanttenetofthismodelisthattheIndsmutationsincre I,fromFlavobacteriumokeanokoites,isamemberoftheunusual,typeIIsclassofrestrictionendonucleasesthatrec-ognizeaspecificDNAsequenceandcleavenonspecificallyashortdistanceawayfromthatsequence(1).Ibindstheco Genetics:Slupskaetal.Table1.ComparisonofmutAandmutCwithmutA,CStrainLac+revertantsper(CC105)108cells,no.+2.3 Proc.Natl.Acad.Sci.USA88(1991)5593Table1.YeaststrainsNuclearMitochondrialStraingenotypegenotypeRef.947aade2ura3-52p09ID41-6aadellysi15161p+15C1036mit-;COXJintron116splicing-defectivepointmutant(trans- Proc.Nat.Acad.Sci.USA68(1971)Torpracticalpurposes,itisconvenienttousethenormal-izedintensitydistributionfunctionIeb(i')N.hv4EEPavl(OavIRabebvN)126(EaV',bV-hv)VIVfo~b(p)N=a'4tt|0D||b)=EEiPavIP4av,,bvNI Cellsrespondtodevelopmentalcuesandenvironmentalstimulibyalteringgeneexpression.Often,transmissionofdifferentupstreamsignalsinvolvescommoncomponentsyetelicitsdistinct(butappropriate)outcomes.Numerousag Proc.Natl.Acad.Sci.USA93(1996)2583multiplexamplificationasdescribedabove.Theonlydiffer-encebetweenthesetwoproceduresisthatsomePprimersusedinMGSmaycontainsingle-basemismatchestothetemplatestoconvertthe

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