APPLICATIONS

-DNApolymerasehavingbothreversetranscriptionandDNAamplificationactivity,excellent foronestepRT-PCRandreal-timequantitativePCR;
-ForhightemperaturereversetranscriptionthatallowshighlyspecificprimingofCDNAsynthesisandovercomesproblemswithsecondarystructure.

DESCRIPTION
TthPlusDNApolymeraseisisolatedfromtheThermusThermophilusstrain.TthPlusDNApolymeraseisasingle92kDapolypeptideshowinga5′-3′exonucleaseactivitybutlacking3′-5′exonucleaseactivity.Itcatalyzesthepolymerizationofnucleotidesintodouble-strandedDNAinthepresenceofMgCl₂.ItsefficiencyhasbeenshownmoreparticularlyonlargeDNAfragmentsupto12kb(usinglamBDaphageDNAasatemplate).TthPlusDNApolymeraseisalsocapableofcatalyzingthepolymerizationofDNAusingaRNAtemplateinthepresenceofMn²⁺.TheABIlityofTthPlusDNApolymerasetoreversetranscribeatelevatedtemperatures(70°C)minimizestheproblemsencounteredwithstrongsecondarystructuresinRNAsincetheyareunstableathigherreactiontemperatures.Highertemperaturesalsoresultinincreasedspecificityofprimerhybridizationandextension.IncoupledRT/PCRassays,TthPlusisabout50-100timesmoreefficientthanregularTaqDNApolymerase.

TthPlusDNApolymeraseisdeliveredwithaone-tube10xbuffer.Bothreactions(RTandPCR)arecarriedoutinthesamebufferandsametubewiththesameenzyme(TthPlus).The10xonetubebufferdoesnotcontainMn(OAc)₂.Mn(OAc)₂isprovidedextraandhavetobeaddedatafinalconcentrationof3.5mM.

 

SIZEandCONCENTRATION

500Units,5units/µl

 

FEATURES

-thermostable;
-TaqDNApolymeraseactivity;
-reversetranscriptaseactivityat70°C.

UNITDEFINITION

Oneunitisdefinedastheamountofenzymethatincorporates10nmolesofdNTPsintoacid-insolubleformin30minutesat72°Cunderthefollowingreactionconditions:25mMTAPSbuffer(Tris-(hydroxymethyl)-methyl-amino-propanesulfonicacid,sodiumsalt)pH9.3(25°C),50mMKCl,2mMMgCl₂,1mMβ-mercaptoethanol,200µMdNTPsand10µgofcalfthymusDNAinafinalreactionvolumeof50µl.

 

STORAGEBUFFER

10mMK-phosphatebufferpH7.0(25°C),100mMNaCl,0.5mMEDTA,1mMDTT,50%glycerol(v/v),0,1mg/mlBSA.

 

STORAGETEMPERATURE

StoreTthPlusDNApolymerasepreferablyat-20°Cinaconstanttemperaturefreezer.

 

COMPONENT

-TthPlusDNApolymerase

-5xOneTubebuffer

-50mMMn(OAc)₂

 

Theoptimalexperimentalconditionsdependonthesystemusedandtheyshouldbeindividuallydetermined.TheMn²⁺concentrationsandtheenzymeamountarethelimitingfactorsforanaccurateresult.TrADItionally5unitsofenzymeandaMn-acetateconcentrationof3.5mMareusedforthereversetranscriptioninafinal50µlreactionvolume.

AdditionalInformation

Clickherefortechnicalinformation

欢迎来到eZYME eENZYME是向科学界提供高品质生物试剂和生物测定试剂盒的制造商和分销商。我们与NIH和CDC社区紧密合作，以确保我们的产品在广泛领域中用于生命科学研究的成功表现。我们努力提供创新且具有成本效益的工具，使您的成功更加轻松。 我们的产品用于学术，生物技术，制药和政府研究实验室的许多不同应用中，包括药物发现，癌症研究，传染病研究，微生物学和分子诊断。 eenzymeeenzyme代理 RANTAI  主营产品：细胞株和菌株、胎牛血清、生化试剂、ELISA试剂盒、抗体和抗原、细胞因子、技术服务、实验耗材和消耗品、仪器设备。 Idtdna 货号：707025 706020 英文名字：Surveyor Mutation Detection KitIdtdna中文名字：基因突变检测试剂盒Idtdna listlabs 以下货号现货 销售eenzymeeenzyme代理 ybiotech品牌工厂 货号：171E 英文名字：Anthrax ToxinsIdtdnalistlabs 中文名字：炭疽毒素listlabs 货号：130A 英文名字：Botulinum Toxinslistlabs中文名字：肉毒毒素listlabs 货号：120 英文名字：Alpha Toxin from Staphylococcus aureuslistlabs中文名字：金黄色葡萄球菌α毒素listlabs 货号：180 英文名字：Pertussis Toxin from B. pertussis, Lyophilized in Bufferlistlabs 中文名字：百日咳毒素listlabs