Description | T7DNAPolymerasecatalyzesDNAsynthesisinthe5'->3'direction.ItisahighlyprocessiveDNApolymeraseallowingcontinuoussynthesisoflongstretchesofDNA.Theenzymealsoexhibitsaverystrong3'->5'exonucleaseactivitytowardssingle-anddouble-strandedDNA. |
Applications | - PurificationofcovalentlyclosedcircularDNAbyremovalofresidualgenomicDNA
- Primerextensionreactionsonlongtemplates
- DNA3'-endlabeling
- Strandextensionsinsite-directedmutagenesis
- Fill-inbluntingof5'-overhangDNA
- SecondstrandsynthesisofCDNA
- InsitudetectionofDNAfragmentationassociatedwithapoptosis
|
Source | T7DNAPolymeraseiscomposedoftwosubunits:an80kDapolypeptide(theproductofgene5ofbacteriophageT7)anda12kDathioredoxinfromthetrxAgeneofE.coli.EachproteinisclonedandoverexpressedseparatelyinE.coli. |
UnitDefinition | Oneunitisdefinedastheamountofenzymethatwillincorporate10nmolofdNTPintoacid-precipitablematerialin30minutesat37°C. |
Components | - T7DNAPolymerase:10,000U/mlin50mMKPO4,0.1mMEDTA,1.0mMDTT,50%glycerol,pH7.0@25°C
- 10XReactionBuffer:200mMTris-HCl,100mMMgCl210mMDTT,pH7.5@25°C
|
QualityControl | Theabsenceofendodeoxyribonucleasesisconfirmedbytheappropriatequalitytest. |
StorageCondition | -20°C |
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