Lucigen/MasterAmp™ ExtraLong PCR 2X Premix 1/MHF925A/5 ml

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.
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.