• Reliablyjoindouble-strandednucleicacidchains.

NxGen Enzymes IconT4DNALigasecatalyzestheformationofaphosphodiesterbondbetweentheterminal5′phosphateand3′hydroxylgroupsofduplexDNAorRNA.TheenzymeefficientlyjoinsbluntandcohesiveendsandrepairssinglestrandednicksinduplexDNA,RNAorDNA/RNAhybrids.

T4DNALigaseVersionSizesLigaseConcentrationProvidedBuffer
LowConcentrationLigase

1,500U
7,500U

2U/µL10XT4DNALigaseBuffer
HighConcentrationRapidKit1,500U
7,500U
10U/µL2XRapidLigationBuffer,
10XT4DNALigaseBuffer

10XT4DNALigaseBufferiscomposedof500mMTris-HCI,100mMMgCl2,50mMdithiothreitol,10mMATP,pH7.6@25°C.

2XRapidLigationBufferiscomposedof132mMTris-HCI,20mMMgCl2,2mMdithiothreitol,2mMATP,15%PEG,pH7.6@25°C.

 

 

Figure1: Purity
ThepurityoftheT4DNAligaseisequalorgreaterthan95%asjudgedbySDS-polyacrylamidegelelectrophoresiswithCoomassiebluestaining. MolecularweightMarkersareinLane1. T4DNAligasewasloadedat5units(Lane2)and10units(Lane3).

 

Figure2: ExonucleaseandEndonucleaseActivityAssay
TheT4DNAligaseisfreeofdetectableofexo-andendonucleaseactivities,asjudgedbyagarosegelelectrophoresisfollowingincubationof10unitsofenzymefor16hoursat37°Cwith1µgofHindIII-digestedλDNA(Lanes3and4)andsupercoiledpUC19DNA(Lanes5and6). MolecularweightmarkersareinLanes1and2.

 

UnitDefinition:OneWeissUnitisdefinedastheamountofenzymerequiredtoconvert1nmolof32P-labeledinorganicpyrophosphateintoNoritadsorbablematerialin20minutesat37°C,usingspecifiedreactionconditions.Note:1WeissUnitisapproximately67cohesiveendunits.

Source:ArecombinantE.colistraincarryingtheclonedT4DNALigasegene.

ORDERINFORMATION

Storagebuffer:T4DNALigaseissuppliedin10mMTris-HCl,50mMKCl,1mMdithiothreitol,0.1mMEDTA,0.1%TritonX-100,50%glycerol,pH7.5@25°C.

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发布于 : 2019-03-27 阅读(0)
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