- GreatTaqPolymeraseperformanceatalowprice.
- Choiceofreactionbuffers,withorwithoutMgCl2.
- Non-proofreADIngPolymerase
FreeSampleAvailable
At$96for1000unitslistprice(quantitydiscountsavailable),EconoTaq’slowpriceiscoupledwithhighqualityandperformance.
QCspecificationsforEconoTaqarerigorous:
- Greaterthan99%purebySDSgelelectrophoresis(seeFigure1).
- NodetectableDNAcontaminationasdeterminedbyPCRusinggenericprimers.
- Nodetectableendonuclease(nicking)activity.Incubationof10UofEconoTaqDNAPolymerasewith1µgofsupercoiledpBR322DNAfor16hoursat70°Cresultsinnodetectableconversiontorelaxedorlinearformsbyagarosegelelectrophoresis.
- Nodetectableexonucleaseactivity.Incubationof10UofEconoTaqDNAPolymerasewith1µgofHindIII-cutlamBDaDNAfor16hoursat70°Cresultsinnosmearingofbandsonagarosegels.
EconoTaqPerformance
AsshowninFigures2and3Lucigen’sEconoTaqDNAPolymeraseperformsaswellas,orbetterthan,moreexpensiveTaqpreparationsfromseveralothersuppliersinroutinePCR.EconoTaqisaseffectiveinDNAamplificationasanotherstandardPCRenzyme,TflDNApolymerase(Figure4).Inthiscase,thebackgroundofnon-specificamplificationwasmuchlowerwithEconoTaq(compare“+”and“–“lanesforEconoTaqandTflinFigure4).EconoTaqDNAPolymerasealsooffershighlot-to-lotreproducibilityandreliABIlity(Figures2and4)
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Figure1.HighpurityofEconoTaqDNAPolymerase(SDSPAGE).Lane1,broadrangemolecularweightMarkers;Lane2,LucigenEconoTaqDNAPolymerase | Figure2.TaqDNApolymerasefromPromegaandNewEnglandBiolabswerecomparedtoLucigen’sEconoTaqDNAPolymerase(2differentlots)inamplifyingtheampicillingene(0.8kb)inapUC19vector.(–),noDNA.(+),DNAadded(40ng).MW,1kbladder. |
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Figure3.EconoTaqvs.AmpliTaq®(AppliedBiosystems)DNApolymeraseingenotyping.AllPCRreactionswereperformedinaRoboCycler96(Stratagene).Hip1genotypingwasperformedusingthefollowingPCRconditions:94°Cfor1min,35cyclesof94°Cfor30sec,62°Cfor60sec,72°Cfor90sec,and72°Cfor7min.Shh,CdoandGas1genotypingwereperformedusingthefollowingPCRconditions:94°Cfor2min,35cyclesof94°Cfor60sec,65°Cfor60sec,72°Cfor90sec,and72°Cfor7min.AllPCRreactionscontainedfinalconcentrationsof1µMofeachprimer,200µMdNTPs,1Xcresolredloadingdye,and1UoftheindicatedTaqpolymerase.SequencesforallPCRprimershavebeenpreviouslypublished(referencesavailable). |
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Figure4.PCRamplificationwasperformedunderstandardconditionsusingthreedifferentlotsofEconoTaqDNAPolymeraseandbuffer,orduplicatereactionswithTflDNApolymeraseandbuffer(Promega).Reactionscontainedprimersspecificforthe16SribosomalRNAgene,withBacillusgenomicDNA(+)ornoDNA(-)asatemplate(1450bpproductexpected). |
PleaseNote:
SomeapplicationsinwhichLucigen'sEconoTaqDNAPolymerasecanbeusedmaybecoveredbypatentsissuedandapplicableintheUnitedStatesandcertainothercountries.Becausepurchaseofthisproductdoesnotincludealicensetoperformanypatentedapplication,usersofthisproductmayberequiredtoobtainapatentlicensedependingupontheparticularapplicationinwhichtheproductisused.ThePCRprocessisthesubjectofEuropeanPatentNos.201,184and200,262ownedbyHoffman-LaRoche.ThosepatentsexpiredonMarch28,2006.ThecorrespondingPCRprocesspatentsintheUnitedStatesexpiredonMarch29,2005.Itisthesoleresponsibilityofthebuyertoensurethatuseoftheproductdoesnotinfringethepatentrightsofthirdparties.
ORDERINFORMATION
EconoTaqDNAPolymeraseisprovidedwithachoiceof10XReactionBufferwithMg++(“withMg++”);or10XReactionBufferwithoutMg++andaseparatetubeof25mMMgCl2(“separateMg++”).PleaseinquireforBulkpricing!ebiomall.com
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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？
