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蚂蚁淘在线 / 品牌中心 / Biomatrica / Biomatrica/PCrbost™ 试剂盒/63301-011/1毫升

Biomatrica/PCrbost™ 试剂盒/63301-011/1毫升

价格
￥5000.00
货号：63301-011
浏览量：127
品牌：Biomatrica
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商品描述

ImproveYourPCRPerformancewithPCRboost.Simply.

  • EnhanceyourmostchallengingPCRsamples
  • SimpletoUse
  • Reducesnumberofcycles
  • UseexistingTaqpolymeraseandPCRprotocol

Overview

EnhancePCRPerformanceforDifficultSamples

PCRBoostisauniquereagentthatenhancesend-pointandreversetranscription-PCRperformancebyimprovingsensitivityandspecificityduringamplificationofgenomicDNAorRNAtemplates.Touse,simplyreplacethewaterinyourPCRwithPCRBoost.That’sall.ItisdesignedtoenhancetheamplificationofDNAwithouttheneedfortimeconsumingoptimizations.PCRBoostsuccessfullyamplifiesthermallydegradedDNAresultinginavisIBLeampliconthatcanbeusedfordownstreamapplicationssuchascloningandsequencing.

ProductData

SuccessfulamplificationofthermallydegradedDNAusingPCRBoost

Figure1:GenomicDNA(gDNA)fromhumanbloodwaspurifiedusingtheQIAamp®DNABloodMiniKit.,and500ngwasexposedto70°Cfor20h.1µl(~50ng)ofthermallydegradedDNAorcontrolDNAstoredat4°CwasusedastemplateforPCRreactions.Thereactionsweresetupusing2.5UTaqDNApolymerase(NEB),3μl10xThermopolreactionbuffer(NEB),0.5µldNTPs(10µMeachnucleotide),0.5µleachofhumanβ-actinforwardandreverseprimers.ThevolumeinthethermallydegradedDNAPCRreactiontubeswasbroughtupeitherinwaterorinPCRBoost.FollowingPCRamplification,10µlofeachPCRreactionwasrunonanagarosegel.A)500nghumangenomicDNAstoredat4°Candthermallydegradedat70°Cfor20hrunonanagarosegel.L:1kbladder.B)50ngofcold-stored(4°C)andthermallydegradedDNAusedastemplateinPCRreactionsamplifyingthehumanβ-actingene.PCRreactionswerebroughtuptovolumeeitherinwaterorPCRBoost.L:ladder.

SerialDilutionUsingPCRBoost

Figure-2:50nghumangenomicDNA(gDNA)wasamplifiedbyPCRusing2.5UTaqDNApolymerase(NEB),3µl10xthermopolreactionbuffer(NEB),0.5µldNTPs(10µMeachnucleotide),0.5µleachhumanβ-actinforward(5’ctacctcatgaagATCCtcacc3’)andβ-actinreverse(5’gtacttgcgctcaggaggagc3’;10µMeach)inafinalvolumeof30µl.ThePCRBoostvolumewasseriallydilutedandreplacedbywater*.Cyclingparameterswere:94°Cfor5minfollowedby40cyclesof94°Cfor15sec,55°Cfor30secand72°Cfor30sec.10µlofeachPCRreactionswasrunona0.8%agarose/ethidiumbromidegel.

IdenticalReactionsinWaterVersusPCRBoost

Figure-3:100,50,20,10or4nggDNAwasusedinPCRreactionswherethereactionwasbroughtuptovolumeineitherwaterorPCRBoost.Sampleswereusedtoamplifythefibroblastgrowthfactor13(FGF13)genebyPCRusing2.5UTaqDNApolymerase(NEB),3µl10xthermopolreactionbuffer(NEB),0.5µldNTPs(10µMeachnucleotide),FGF13forward(5’gAATgttaacaacatgctggc3’)andFGF13reverse(5’agaagctttaccaatgttttcca3’)(kindgiftofDr.D.Cohn)inafinalvolumeof30µl*.Cyclingparameterswere:94°Cfor5minfollowedby40cyclesof94°Cfor15sec,55°Cfor30secand72°Cfor30sec.10µlofeachPCRreactionwasrunona0.8%agarose/ethidiumbromidegel.

ComparisonofPCRBoosttoOtherPCREnhancers

Figure-4:PCRenhancersfrom3differentcompaniesweretestedalongsidePCRBoostandcomparedtoanon-enhancedwatercontrol*using1nggDNA(conditionsusedwerethesameasFigure2above).

UseofPCRBoostwithThreeDifferentTaqPolymerases

Figure-5:TaqDNApolymerasesfrom3differentsuppliersweretestedalongsidePCRBoostusing10nggDNA*(conditionsusedwerethesameasFigure2above).

TestingofPCRBoostinRT-PCR

Figure-6:RT-PCRreactionsweresetupusingthegenespecificreverseprimerforthesingle-copyRNasePgene(5’agaccatcctggctaacacg3’).SmallamountsoftotalRNA(100ng)extractedfrom293cells(humanadenocarcinomacellline)wereusedforfirststrandCDNAgenerationusingmanufacturer’sinstructionsfortheAffinityScript™(Stratagene/AgilentTechnologies)RT-PCRkit.ReactionsweresetupusingeitherwaterorreplacingthewatercomponentwithPCRBoost.1µlofthecDNAwasthenusedinasubsequentsecondstrandPCRreactionusingforward(5’ttcactgcttcatgcctacg3’)andreverseprimersfortheRNasePgene(conditionsusedwerethesameinFigure2above).ReactionsweresetupusingeitherwaterorreplacingthewatercomponentwithPCRBoost.

*PCRreactionsweresetupusingcocktailsofcommonelements.

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Specifications

  • Productformat: 1mL
  • Storagetime: 6months
  • Storagetemperature: 15–25°C
  • Downstreamapplications:plasmidpurification,bacterialpropagation,glycerolstockpreparation

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PCRboost
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目前，生物材料，如核酸，蛋白质，细胞，病毒和疫苗等需要在寒冷的环境中保存和运输（4 - 20ºC,-80ºC液氮）才能保持样品的完整性。多次冻融或长时间暴露于过高的温度环境下会降低生物材料稳定性。但在持续的寒冷的环境，就需要不断的能源和资源供应。因此，运输和储存生物样品往往涉及到大量的费用和复杂的物流。尽管精心注意保持样品的完整性，但直到现在运输过程也难以控制的所有环境变量。biomatrica提供了一个创新性方案来解决对冷链物流的依赖。biomatrica已经开发了samplematrix®稳定专有的核心技术能保护生物材料在室温下无降解。该samplematrix技术是由极端微生物在非常干燥的环境中长期生存的生物相结合的设计，通过利用这些独特的特点，biomatrica产品使研究人员在学术界，药物开发，和分子诊断的生物样品在运输和储存很长一段时间后保持样品稳定性，并且可以降低成本。实验样品的稳定性保证了实验的可重复性，从而保证实验的可靠性，效率，节省时间等，这样就大大降低了成本，这样的技术是以往样本低温保存的理想替代品。这项技术可用于生物技术、诊断、制药工业等成千上万的即用型产物，也可直接用于来自基因组图谱的数十亿样本，以及法律和军事中用于个人药检和刑侦分析的存档样本。