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Ligatesingle-strandedadenylatedDNAorRNAoligostoRNAtargetsforavarietyofapplications.
Ligatesingle-strandedadenylatedDNAorRNAoligostoRNAtargets- UsethisenzymetoprepareCDNAlibrariesforsmall-RNAtranscriptomeanalysissuchasRNA-Seq
- PerformlinkerligationformiRNAcloningapplications
T4RNALigase2,DeletionMutant,alsoknownasT4Rnl2(1-249),isusedtoligatesingle-strandedadenylatedDNAorRNAoligonucleotidestosmallRNAsforcloningornext-generationRNAsequencing.Thepreadenylated5´endsofDNAorRNAareligatedtothe3´endsofRNA.Unlikethefull-lengthenzyme,T4Rnl2(1-249)isunabletoadenylatethe5´endofthesubstrateinthepresenceofATP.However,itcanuseapreactivateddonor(App-DNAorApp-RNA)andjoinittothe3´endofanacceptor;thus,performingtheligationreactionintheabsenceofATPwhichpreventscircularizationandotherundesirablebimolecularreactions.
Theenzymeisavailablein2,000and10,000-Unitsizesataconcentrationof200U/µl.Theenzymeissuppliedwitha10XReactionBuffer.
UnitDefinition:Oneunitistheamountofenzymerequiredtogive50%ligationofa22-merRNAtothepreadenylatedendofa17-merDNAwhenbotholigosareannealedtoacomplementary39-merDNAstrandin30minutesat37°Cunderstandardassayconditions.
StorageBuffer:50%glycerolcontaining50mMTris-HCl(pH7.5),0.1MNaCl,0.1mMEDTA,1mMdithiothreitol(DTT),and0.1%Triton®X-100.
T4RNALigase2,DeletionMutant,10XReactionBuffer:500mMTris-HCl(pH7.5),20mMMgCl2,and10mMDTT.
ActivityAssay:Theunitdefinitionassayisperformedinareactioncontaining50mMTris-HCl(pH7.5),2mMMgCl2,1mMDTTand0.4µgofequimolarmixof22-mer,17-merand39-meroligonucleotides,andvaryingamountsofenzyme.
QualityControl:T4RNALigase2,DeletionMutant,isfreeofdetectableDNAexo-andendonuclease,andRNaseactivities.
ORDERINFORMATION
T4RNALigase2,DeletionMutantisprovidedat200U/μl.
Contents:T4RNALigase2DeletionMutant,10XReactionBuffer.
美国Lucigen公司，自1998年成立至今，一直致力于生命科学领域相关科研产品的研究与开发，在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品，包括：CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等。Lucigen公司凭借其独到的产品技术，过硬的产品质量，良好的产品服务赢得了全球广大用户的信赖。
