OptimizePCRamplificationoflongDNAampliconsupto40kb
Fast,simpleoptimizationstartingwiththecompletekit- EasilyamplifyDNAtargetsupto40kblong
- ReducePCR-inducederrorsintheselongampliconswiththishighfidelityDNApolymerasemix
TheMasterAmp™Extra-LongPCRKitisacompletesystemforsuccessfulandaccurateone-stepamplificationofDNAsequencesupto~40kb.ThekitcontainstheMasterAmp™Extra-LongDNAPolymeraseMix,aswellasnineMasterAmpExtra-LongPCR2XPreMixeswithdNTPs,buffer,andvaryingamountsofMgCl2andtheMasterAmpPCREnhancerwithBetaine.*
Werecommendfirstusingthecompletekit(Cat.No.MHF9220)totestthedifferentMasterMixExtra-Long2XPreMixes(Step1,Fig.1)witheachnewtemplateandprimersettoidentifythebestperformingone.OnePreMixwillamplifyyourlongtemplateandgiveconsistentresultsinallsubsequentamplifications(Step2,Fig.1).OncetheoptimalPreMixisidentifiedcontinuetousethatcombinationofMasterAmpExtra-Long2XPreMixandDNAPolymeraseforallsubsequentamplificationsusingthattemplateDNAandprimerset.
| Figure1.TwostepstoreliableandconsistentPCRresults. | |
| STEP1.FindtheoptimalMasterAmp™Extra-LongPCRPreMix.PerformPCRwithyourtemplate,primers,andtheninePreMixes. | STEP2.UsethesameMasterAmp™Extra-LongPCRPreMixtoobtainreliable,consistentPCRofthesamesequence. |
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| A.Amplificationofa20-kbregionoflamBDaDNAusingMasterAmp™Extra-LongPCR2XPreMixes(1-9).MasterAmp™Extra-LongPCRPreMix4producedoptimalresults. | B.SubsequentPCRofa20-kblambdaDNAregionwiththesameprimerpairandMasterAmp™PreMix4gaveconsistentresults,usingtheoptimalPreMixdeterminedinStep1. |
 | Figure2.Extra-longamplificationoflambdaDNAusingtheMasterAmp™Extra-LongPCRKit.PCRconditionswereoptimizedandMasterAmpPremix4wasselectedforPCRof1ngoflambdaDNAineachsample.Resultswereanalyzedona0.5%agarosegel.Ampliconsizeswere30kb(lane1),35kb(lane2)and40kb(lane3).LaneM,5-KbDNAladder. |
*Coveredbyissuedand/orpendingpatents.
MasterAmp™Extra-LongPCRKit;Purchaseofthisproductincludesanimmunityfromsuitunderpatentsspecifiedintheproductinserttouseonlytheamountpurchasedforthepurchaser"sowninternalresearch.Nootherpatentrights(suchas5´NucleaseProcesspatentrights)areconveyedexpressly,byimplication,orbyestoppel.FurtherinformationonpurchasinglicensesmaybeobtainedbycontactingtheDirectorofLicensing,AppliedBiosystems,850LincolnCentreDrive,FosterCity,California94404,USA.
