Description
MethylampHotTaqDNAPolymeraseisachemicallymodifiedMethylampTaqDNAPolymerase.Atambienttemperaturesitisinactive,havingnopolymerizationactivity.MethylampHotTaqDNAPolymeraseisactivatedbya15minincubationstepat95°C.Thispreventsextentensionofnon-specificallyannealedprimersandprimer-dimersformedatlowtemperaturesduringPCRsetup.Theenzymehas5’→3’polymerizationdependentexonucleasereplacementactivitybutlacks3’→5’exonucleaseactivity.

Concentration
5units/µl(oneunitisdefinedastheamountofenzymerequiredtocatalyzetheincorporationof10nmolofdNTPintoanacid-insolubleformin30minat74°C).

Source
PurifiedfromanE.colistraincarryinganoverproducingplasmidcontainingamodifiedgeneofThermusaquaticusDNAPolymerase.

Storage&DilutionBuffer
50%glycerol(v/v),20mMTris-HClpH8.7at25°C,100mMKCl,0.1mMEDTAandstABIlizers.

Composition

• MethylampHotTaqDNAPolymerase 
• 10xReactionBufferB1(Mg2+,detergentfree):Tris-HCland(NH4)2SO4 
• 10xReactionBufferB2(Mg2+free):Tris-HCl,(NH4)2SO4anddetergent 
• 25mMMgCl2
• 10xEnhancer
  Additivethatfacilitatesamplificationofdifficulttemplates(e.g.GC-richDNAtemplates). Enhancershouldbeusedatadefinedworkingconcentration(1x,2xor3xsolution). 

Applications

• HotStartPCR 
• Primerextension 
• TAcloning

StorageConditions
Temporarystorageforupto1monthatroomtemperaturehasnodetrimentaleffectsonthequalityofthisreagent.However,routinestorageat-20ºCisstronglyrecommended.

Fig.1. Amplificationof~500bpand~600bptargetDNAfragmentsusingMethylampHotTaqDNAPolymerase.

1-3–MethylampHotTaqDNAPolymerase

M–EpiQuik1KbDNALadder