Transcription of DNA sequencing with chain-terminating inhibitors
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, , ,December1977 BiochemistryDNAsequencingwithchain-termi natinginhibitors(DNApolymerase/nucleotid esequences/bacteriophage4X174) , , ,CambridgeCB22QH, ,October3, "plusandminus"method[Sanger, , (1975) ,441-4481butmakesuseofthe2',3'-dideoxyan darabinonu-cleosideanaloguesofthenormald eoxynucleosidetriphosphates, "plusandminus"method(1)isarelativelyrapi dandsimpletechniquethathasmadepossibleth edeterminationofthesequenceofthegenomeof bacteriophage4X174(2). ,neitherthe"plus"northe"minus"methodisco mpletelyaccurate,andinordertoestablishas e-quencebothmustbeusedtogether, ( ,inpress)hasrecentlydevelopedathirdmetho d,involvingribo-substi-tution,whichhasce rtainadvantagesovertheplusandminusmethod , (3), ,butitrequiresastrandseparationorequival entfrac-tionationofeachrestrictionenzyme fragmentstudied, , (4)showedthattheinhibitoryactivityof2',3 '-dideoxythymidinetriphosphate(ddTTP)onD NApolymeraseIdependsonitsbeingincorpo-ra tedintothegrowingoligonucleotidechainint heplaceofthymidylicacid(dT).]
Proc. Natl. Acad.Sci. USA74(1977) primerandthe newlysynthesized DNA.Thisis themostsimple andrapidprocedure, requiringonlyapreliminary annealing oftemplateandprimer ...
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