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当前位置: 首页 > 产品中心 > Reverse Transcriptase Enzymes > Lucigen/EpiScript™ RNase H- Reverse Transcriptase/ERT12910K/10,000 Units
公司介绍
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
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Lucigen/EpiScript™ RNase H- Reverse Transcriptase/ERT12910K/10,000 Units

Lucigen/EpiScript™ RNase H- Reverse Transcriptase/ERT12910K/10,000 Units
  • Lucigen/EpiScript™ RNase H- Reverse Transcriptase/ERT12910K/10,000 Units
商品介绍

Anoptimizedreversetranscriptasesystemfortheproductionoffull-lengthCDNA.

  • Formerly from EpicentreProducefull-lengthcDNAwithRNAseH-mutant
  • Achievehigherspecificityatelevatedtemperatures,upto55°C

EpiScript™RnaseH-ReverseTranscriptase(EpiScriptRT),analternativetoSuperScript®IIReverseTranscriptase,isarecombinantMMLVreversetranscriptasewithgreatlyreducedRNaseHactivity.Itishighlyefficientatproducingfull-lengthcDNAfromlongRNAtemplates.EpiScriptRTiscapableofproducingcDNAfromaslittleas50pgoftotalRNAforreal-timeRT-PCR(qRT-PCR)analysisandotherapplications.

Applications

  • First-strandcDNAsynthesisforsubsequentPCRorreal-timePCR.

Benefits

  • RecombinantMMLVreversetranscriptasewithgreatlyreducedRNaseHactivity
  • Activeattemperaturesupto55°C
  • Highlyefficientatproducingfull-lengthcDNAfromaslittleas50pgoftotalRNA
  • BestvalueinanRNaseH-ReverseTranscriptase

Storage:Storeonlyat-20°Cinafreezerwithoutadefrostcycle.

StorageBuffer:EpiScriptRTissuppliedina50%glycerolsolutioncontaining50mMTris-HCl(pH7.5),100mMsodiumchloride,1mMDTT,0.1mMEDTA,and0.1%Triton®X-100.

UnitDefinition:OneunitofEpiScriptRTcatalyzestheincorporationof1nmolofdTTPintoacid-insolublematerialin10minutesat37°Cusingsaturatingamountsofoligo(dT)-primedpoly(rA)astemplate.

ContaminatingActivityAssays:EpiScriptRTisfreeofdetectableexonuclease,endonuclease,andRNaseactivities.

Figure 1

Figure1.EpiScript™ReverseTranscriptaseperformedequallyorbetterthancomparablereversetranscriptasesfromothervendors.First-strandsynthesisreactionswereassembledaccordingtomanufacturer´sspecifications.InputRNAwas1µgofJurkattotalRNA(Ambion®).Reactionswereprimedusing50ngofpoly-T(16-18)DNA.2ndstrandqPCRwasperformedusingBio-RadiQSYBRmastermixandgene-specificprimersthatyielded250-350bpamplicons.Reactionswererepeated4-fold.PGDF-R(PlateletDerivedGrowthFactorReceptor),TNF(TumorNecrosisFactor),IL-1b(Interleukin-1beta),IL-2(Interleukin2).ImagecourtesyofMatthewKellinger,Illumina®Inc.

Figure 2AFigure 2BFigure2(clicktoenlarge).EpiScript™ReverseTranscriptaseproducessimiliartranscriptcoverageindependentoftranscriptlength.EpiScriptwasusedtoprimefirststrandcDNAeitherfromtotalRNAusingoligo-dT(leftpanel)orpolyA+selectedRNAwithrandomhexamers(rightpanel).ThecDNAwasconvertedintoIllumina®-compatIBLelibrariesandsequenced.Thereadswerealignedtotranscriptsbaseduponvariouslengthclassesandreaddensityplottedrelativetothepercentdistancefromthe5´endofthetranscripts;0%referstothe5´endand100%isthe3´end.Asexpected,oligo-dTprimingresultsinamorepronounced3´biasthanrandompriming.

Figure 3Figure3.UseofEpiScript®ReverseTranscriptaseforfirststrandcDNAresultsindetectionofsimilartranscriptcategoriesindependentofinputamount.EpiScriptRTwasusedtorandomprime5ngor50pgofhumantotalRNAandthecDNAwasconvertedintolibrariesforIllumina®sequencing.ReadswerealignedusingTophatandannotatedwithCufflinksandthepercentageofeachmajorcategoryisvisualizedasapiechart.Equivalentresultsareobservedforboth5ngand50pgofinputRNA.

ORDERINFORMATION

Contents:EpiScriptRNaseH-ReverseTranscriptase,10XReactionBuffer,100mMDTT.
品牌介绍

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。


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