DNASEQUENCING

LUCIADHIANTIKAWITASARI

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Genomicsequencingtimeline

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 DNApolymerasesrequire
bothaprimer(ashort
oligonucleotide strand),to
whichnucleotidesare
added,andatemplate
strandtoguideselectionof
eachnewnucleotide.
Incells,the3hydroxyl
groupoftheprimerreacts
withanincoming
deoxynucleoside
triphosphate (dNTP)toform
anewphosphodiester bond.

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 Dideoxynucleoside triphosphate (ddNTP)analogs

TheSangersequencing
procedureuses
dideoxynucleoside
triphosphate (ddNTP)analogs
tointerruptDNAsynthesis.
(TheSangermethodisalso
knownasthedideoxy method.)
WhenaddNTP isinsertedin
placeofadNTP,strand
elongationishaltedafterthe
analogisadded,becauseit
lacksthe3hydroxylgroup
neededforthenextstep.
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 SEKUENSINGDNA
METODESANGER

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 DNAsequencingisreadilyautomatedbya
variationofSangerssequencingmethodin
whichthedideoxynucleotides usedforeach
reactionarelabeledwithadifferentlycolored
fluorescenttag.
ThistechnologyallowsDNAsequences
containingthousandsofnucleotidestobe
determinedinafewhours

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