OptimizePCRreactionsthatusedifferentenzymesthantheFailSafe™System(e.g.Taq,Pfu,andTth).
Fast,simpleoptimization- ExtremelyhighsensitivityandspecificityusingthePCREnhancerTechnology
- MasterAmpPCRPreMixescontainallcomponentsneededforsuccessfulPCR
- GeneratesPCRproductssuitableforbothTAcloningandblunt-endcloning
- UsefortheoptimizationofPCRreactionsusingavarietyofdifferentenzymesincludingTaq,Pfu,Tth,Tflpolymerases
Applications
Forrapid,dependableoptimizationofPCR,weencourageyoutotrytheFailSafe™PCRSystemtoobtainthebestPCRresultswithanytemplateupto~20kb.However,ifyouprefertouseanotherenzyme,EpicentrealsohastwoMasterAmp™PCROptimizationKitsthatwillhelpyoutoobtainthebestpossIBLePCRresultswiththermostableenzymesfrequentlyusedinPCR,suchasTaq,Pfu,Tth,TflDNApolymerases.TheseMasterAmpPCROptimizationKitscontainaseriesof12MasterAmpPCR2XPreMixesthathavebeenmeticulouslytestedandrepresentacompleterangeofPCRconditionsfrequentlyencountered.EachMasterAmp2XPreMixcontainsallcomponentsneededforPCRexceptfortheprimers,thetemplate,andthePCRenzyme. First,usetheMasterAmp™PCROptimizationKit(Cat.No.MO7201)toperformPCRwithyourtemplate,primers,andeachofthe12MasterAmpPreMixes.SelectthePreMixthatgivesoptimalresults.Then,usethesameMasterAmpPCRPreMixtoobtainreliable,consistentPCRusngthesameDNAtarget/primerset. |  | STEP1.FindtheoptimalMasterAmpPCRPreMix.DoPCRwithyourtemplate,primersandPCRenzymeinthe12PreMixes. |
A.A75%-GC-richApoEgeneregionwasamplifiedintotalhumangenomicDNAusingPreMixesA-L.MasterAmpPCRPreMixKproducedoptimalresultsinStep1. | ||
 | STEP2.UsethesameMasterAmpPCRPreMixtoobtainreliable,consistentPCRofthesamesequence. | |
| B.SubsequentPCRoftheApoEgenewiththesameprimerpairandMasterAmp™PreMixKgaveconsistentresults.SubsequentPCRresultsareconsistentandreliableusingtheoptimalPreMixinStep1. |
MasterAmpPCROptimizationKits;MasterAmpTaqDNAPolymerase;MasterAmpTflDNAPolymerase.Purchaseofthisproductincludesanimmunityfromsuitunderpatentsspecifiedintheproductinserttouseonlytheamountpurchasedforthepurchaser'sowninternalresearch.Nootherpatentrights(suchas5´NucleaseProcesspatentrights)areconveyedexpressly,byimplication,orbyestoppel.FurtherinformationonpurchasinglicensesmaybeobtainedbycontactingtheDirectorofLicensing,AppliedBiosystems,850LincolnCentreDrive,FosterCity,California94404,USA.
Ep
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
半定量反转录－聚合酶链反应（semi-quantitative reverse transcription and polymerase Chain reaction ,SqRT-PCR）是近年来常用的一种简捷、特异的定量RNA测定方法，通过mRNA反转录成cDNA，再进行PCR扩增，并测定PCR产物的数量，可以推测样品中特异mRNA的相对数量。
定量RT-PCR（quantitative reverse transcription and polymerase Chain reaction ,qRT-PCR）是在用一步法或两步法，在PCR反应体系中加入荧光基团，利用荧光信号积累实时监测整个PCR进程，最后通过标准曲线对未知模板进行定量分析的方法。
半定量RT-PCR需要跑电泳，根据条带亮度的强弱来判断模板拷贝数的高低或者是表达量的高低，而定量RT-PCR则无需电泳可以实时监测整个PCR的全程并且由给出的Ct值及Standard Curve来判断gene拷贝数的高低。
在另外的一组管子中，加入已知拷贝数的DNA同时扩增，每个管子中加入的数量是不同的，但是从最小数目到最大数目的这个范围涵盖了样本中DNA拷贝数。
PCR反应完成后，把已知拷贝数DNA的量和PCR荧光Ct值制成标准曲线，再把待测样本的CT值和该标准曲线比对，就可以得到样本中的起始拷贝数了
有大神做过人外周血全血miRNA的提取及后续PCR实验的吗？目前课题实验遇到瓶颈，求助大神！！！是否miRNA的PCR验证在全血做不出？必须要用血浆或者血清？目前血浆、血清也有部分样本。。。。有大神指条明路吗？
