MCLAB/2X MCAmp Library Amplification Master Mix/LIBA250/1 Ea

Description
The2XMCAmplibraryamplificationmastermixhasbeenspeciallyformulatedtoprovideaccurateandrobustamplificationofNGS(next-generationsequencing)libraryfragments.Themastermixutilizesaproprietarycombinationofourultra-fast,high-fidelityDNApolymerases.Thepolymerasesallpossess3'->5'exonuclease(proofreading)activitywithaprocessivitytentimesgreaterthangenericDNApolymerases.ThereactionbufferhasbeenoptimizedtoproviderobustamplificationoflibraryfragmentsevenwithlowinputandGCorATrichsequences.Theresultingamplificationshaverobustyieldwithhigh-fidelity,lowPCRduplication,andlowbiaswhichproducesgreaterandmoreuniformcoverageespeciallyinchallenginggenomicregions.
ForIllumina^®platforms,anoptimized10XPrimerSettargetingtheP5andP7sitesisalsoavailable.

Producthighlights

• High-fidelity: ThemastermixutilizesDNApolymerasesthatallpossess3'->5'exonuclease(proof-reading)activityforaccuratenucleotideincorporationsintoyourfinalproducts.
• High-efficiency: Thepolymerasesinthemastermixhaveultra-highprocessivitysothatlesscyclesarerequiredforsequencereadylibraries.
• LowbiasandPCRduplication: OptimizedbufferformulationtominimizeamplificationbiasandPCRduplication.Thisresultsinagreaternumberofreadswithmoreuniformcoverage,helpingtominimizecostandtime.

Recommendedstoragecondition:-20°C
HeatInactivation: No

UnitAssayConditions:
25mMTAPS-HCl(pH9.3@25°C),50mMKCl,2mMMgCl₂,1mMβ-mercaptoethanol,200μMdNTPsincluding[3H]-dTTPand400μg/mlactivatedCalfThymusDNA.

UnitDefinition
Oneunitisdefinedastheamountofenzymethatwillincorporate10nmolofdNTPintoacidinsolublematerialin30minutesat74°C.

Figure:LibrarypreparationandamplificationresultsfromvaryinginputquantitiesofshearedhumangenomicDNA.Librarypreparationandsizeselection(~350bp)wascarriedoutusingMCLAB'sTopomizeDNALibraryPrepKit.Allamplificationswerecarriedouttoeightcycles.
A.Agarosegelimageafterlibraryamplificationandcleanupshowingstablesizedistribution.M:100bpMarker.Lane1:100nginputDNA.Lane2:50nginputDNA.Lane3:25nginputDNA.B.AverageDNAyieldofcorrespondinggelagarosesamplesdeterminebyqPCRusingMCNexttmSYBR^®FastqPCRLibraryQuantifiactionKit.