Description:
T7ExonucleaseissimilartoLambdaExonucleaseinthatitcatalyzesthestepwisehydrolysisofduplexDNAfromthe5'-termini,liberating5'-mononucleotides.However,unlikeLambdaExonuclease,theenzymehaslowprocessivityanditwillremoveboth5'-hydroxyland5'-phosphoryltermini.T7ExonucleasehydrolyzesduplexDNAnon-processivelyinthe5'->3'directionfromboth5'-phosphorylor5'-hydroxylnucleotidesbyliberatingoligonucleotides,aswellasmononucleotides,untilabout50%oftheDNAisacidsoluble.

Source:
PurifiedfromanE.colistraincontainingaTYB₁₂inteinfusion.

StorageConditions
10mMTris-HCl
5mMDTT
0.1mMEDTA
50%Glycerol
pH8.0@25°C

Application:
-Controlledstepwisedigestionofdouble-strandedDNAfromthe5'-termini.
-GeneratingssDNAtemplatesforsequencingviathechain-terminationmethod.

Suppliedwith:
10XT7ExonucleaseBuffer

10XT7ExonucleaseBuffer:
500mMPotassiumAcetate
200mMTris-acetate
100mMMagnesiumAcetate
10mMDTT
pH7.9@25°C

UnitDefinition:
Oneunitistheamountofenzymerequiredtorelease1nmolofacidsolublenucleotidein15minat37°Cunderstandardassayconditions.

RecommendedStorageCondition:-20°C

References:
1.Kerr,C.andSadowski,P.D.(1972)J.Biol.Chem.247,311-318.
2.Thomas,K.R.andOlivera,B.M.(1978)J.Biol.Chem.253,424-429.
3.Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.andStruhl,K.,(1987)CurrentProtocolsinMolecularBIOLOGy(JohnWileyandSons,Inc.
4.Shon,M.,Germino,J.andBastia,D.(1982)J.Biol.Chem.257,13823-13827.
5.Nikiforov,T.T.,Rendle,R.B.,Goelet,P.,Rogers,Y.H.,Kotewicz,
6.M.L.,Anderson,S.,Trainor,G.L.andKnapp,M.R.(1994)Nucl.AcidsRes22,(20),4167-4175.
7.Kornberg,A.andBaker,T.(1991)DNAReplication,SecondEdition,591.