Description:
LongAmpHotStartTaq2XMasterMixcontainsauniqueblendofaptamer-basedHotStartTaqandDeepVentR™DNAPolymerases.Theaptamer-basedinhibitorbindsreversIBLytotheenzyme,inhibitingpolymeraseactivityattemperaturesbelow45°C,butreleasestheenzymeduringnormalPCRcyclingconditions.ThispermitsassemblyofPCRreactionsatroomtemperature.TheLongAmp®HotStartTaq2XMasterMixdoesnotrequireaseparatehightemperatureincubationsteptoactivatetheenzyme.The3´→5´exonucleaseactivityofDeepVentRDNAPolymeraseincreasesthefidelityandrobustamplificationofHotStartTaqDNAPolymerase(1).LongAmp®HotStartTaqDNAPolymeraseofferstwo-foldhigherfidelitythanHotStartTaqDNAPolymerasealone.Theconvenientmastermixformulationissuppliedata2XconcentrationandcontainsdNTPsandMg++,requiringonlytheadditionofprimersandDNAtemplateforrobustamplification.AwiderangeofPCRproductscanbegenerated;upto30kbfromlamBDaorhumangenomicDNA.
Amplificationofa8kbampliconfromvaryingamountsofJurkatgenomicDNAusingLongAmpHotStartTaq2XMasterMix.Startingtemplateamountsareindicatedbelowthegel.MarkerMistheNEB2-LogDNALadder(NEB#N3200).ProductSource
AnE.colistrainthatcarriestheTaqDNAPolymerasegenefromThermusaquaticusYT-1andanE.colistrainthatcarriestheDeepVentRDNAPolymerasegenefromPyrococcusspeciesGB-D.
UnitDefinition:
Oneunitisdefinedastheamountofenzymethatwillincorporate10nmolofdNTPintoacidinsolublematerialin30minutesat75°C.
UnitAssayConditions:
1XThermoPol™ReactionBuffer,200µMdNTPsincluding[3H]-dTTPand200 µg/mlactivatedCalfThymusDNA.
ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration |
LongAmp® TaqReactionBufferPack | -20 | 5X |
Notes:
ProductspecificationsforindividualcomponentsintheLongAmpHotStartTaq 2XMasterMixareavailableseparately.