MMLV(MoloneyMurineLeukemiaVirus)ReverseTranscriptaseisaDNApolymerasethatsynthesizesacomplementaryDNAstrandsfromsingle-strandedRNA,DNA,oranRNA-DNAhybridasatemplate.ThisrecombinantenzymewaspurifiedfromE.coli,whichcarriedamodifiedMMLV-RTgene.ComparedtoAMVReverseTranscriptase,thisenzymehasamuchweaker5"-3"ribonucleaseHactivity,whichallowsthesyntesisoflongercDNAs(>7kb).
RecombinantE.colistrain.
SterileFilteredclearsolution(200U/µl).
50mMTris-HCl,0.1MNaCl,0.1%TritonX-100,2mMDTT,0.1mMEDTAand50%glycerol.
Stablefor5daysat10°C,forlongerperiodoftimestoreat-20°C.
Pleasepreventfreeze-thawcycles.
Oneunitisdefinedastheamountofenzymerequiredtocatalyzetheincorporationof1nmolofdeoxyribonucleotideintoacid-insolubleformsin10minutesat37oC,usingpoly(A)-oligo(dT)12-18asthetemplate-primer.Standard
CDNASynthesisConditions50mMTris-HCl(pH8.3),75mMKCl,3mMMgCl2,10mMDTT,1.0mMeachdATP,dGTP,dCTP,anddTTP,0.2mgradomhexamer,1-5mgRNA,200unitsM-MLVRT.Thereactionvolumewas20mlandtheincubationwas45minat42oC.
250mMTris-HCl(pH8.3),375mMKCl,15mMMgCl2,and50mMDTT.
Prospec"sproductsarefurnishedforLABORATORYRESEARCHUSEONLY.Theproductmaynotbeusedasdrugs,agriculturalorpesticidalproducts,foodadditivesorhouseholdchemicals.