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蚂蚁淘在线 / 品牌中心 / Excellgen / Excellgen/ExcellScript M-MLV (MMLV) Reverse Transcriptase RNase H minus/EG-1061/20,000 Units

Excellgen/ExcellScript M-MLV (MMLV) Reverse Transcriptase RNase H minus/EG-1061/20,000 Units

价格
￥1000.00
货号：EG-1061-20,000Units
浏览量：127
品牌：Excellgen
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商品描述
Description

ExCellscriptM-MuLVReverseTranscriptaseRNaseHminusisanRNA-dependentDNApolymerasewithnodetectableRNaseHactivity.ExcellScriptcanbeusedtogeneratefirst-strandCDNAfrompolyAmRNAortotalRNAforuseindownstreamapplicationssuchasRT-PCR,cDNAcloningorlibraryconstructionforRNA-Seq.PointmutationsintheRNaseHdomainincreasetheThermostABIlityoftheenzymeandsupportgreatercDNAyieldoffull-lengthtranscriptsthanwildtypeM-MLVReverseTranscriptase

Applications
  • FirststrandcDNAsynthesisforRT-PCRandreal-timeRT-PCR
  • SynthesisofcDNAforcloningandexpression
  • GenerationoflabeledcDNAprobesformicroarrays
  • DNAlabeling
  • AnalysisofRNAbyprimerextension
SourceArecombinantE.colistraincarryinganengineeredMoloney-MurineLeukemiaVirusReverseTranscriptasegene
UnitDefinition1unitisdefinedastheamountofenzymerequiredtoincorporate1nmolofdTTPintoacidinsolublematerialin10minutesat37°Cusingpolyr(A)/oligo(dT)asasubstrate.
Components
  • M-MuLVReverseTranscriptase:200units/µLin50mMTris-HCl,150mMNaCl,0.1mMEDTA,1mMDTT,0.1%NP-40,50%glycerol,pH7.6@25°C
  • 10XReactionBuffer:500mMTris-HCl,750mMKCl,30mMMgCl2,100mMDTTpH8.3@25°C
StorageCondition-20°C
Protocol
  1. PrimerAnnealing:CombinethefollowinginanRNase-freereactionvessel:
AmountDescriptionFinalConcentration
1µL25mMdNTPSolution(N2050L2.0mM
XµL1ng-2µgTotalRNA-or-
XµL5-500ngmRNA(polyAselected)
1µLOligo(dT)12-18(500µg/ml)-or-40µg/mL
1µLRandomPrimers(125µg/ml)-or10µg/mL
1µLGSPPrimer(2pmol)165µM
XµLSterileWaterN/A
10µLTotalVolume
  1. Heatreactionfor5minutesat65°C.Spinbriefly(5sec)topulldowncondensateandplaceimmediatelyonice.
  2. Add1µL10XM-MuLVRTBufferandDNase-freewatertoafinalvolumeof10µLperreaction.
  3. Incubate:
    • IfusingOligo(dT)orGSPprimers:2minutes@42°C
    • IfusingRandomprimers:2minutes@25°C
  4. Add1µL(200units)M-MuLVReverseTranscriptaseandmixbygentlypipettingsample.(Note:ifusingrandomprimers,pre-incubatereaction@25°Cfor10minutes).
  5. Incubateat42°Cfor45-60minutes.
  6. Inactivateenzymeat85°Cfor10minutes.
  7. Storeproductsat-20°Corproceedtonextstep.
Excellgen蛋白表达产品名称-目录 ＃价钱293Expresso™转染试剂EG-1089$ 98.00定制抗体生产和免疫测定开发EG-36从培养基中纯化IgEEG-1076-1$ 750.00MembraneExpress TM功能膜蛋白的病毒样颗粒EG-1038蛋白质表达和纯化服务EG-1076-8$ 1,500.00小规模表达测试和纯化方法开发EG-1081-3$ 1,250.00使用HEK 293细胞的瞬时蛋白质生产EG-1080$ 2,900.00  $ 1,950.00-$ 950.00使用细菌表达系统进行蛋白表达和纯化EG-1076$ 2,170.00使用杆状病毒表达系统进行蛋白质表达和纯化EG-1078$ 4,500.00使用Akta FPLC系统的定制蛋白质纯化服务EG-1082$ 1,000.00使用稳定的哺乳动物细胞系表达蛋白质EG-1081$ 9,500.00