Description: ThePyrophosphatase,inorganic(PPase)catalyzesthehydrolysisofinorganicpyrophosphatetotwoorthophosphates.Theenzymerequiresadivalentmetalcation,withMg2+conferringthehighestactivity.
Application: -HighyieldsynthesisofRNAbyinvitrotranscription(1,2) -DNApolymerizationreactions:preventingaccumulationofpyrophosphate(3,4) -RemovalofcontaminantPPiinreagentsusedforSNPgenotypingbymethodsbasedonthedetectionofpyrophosphate(5)
Source: E.colicellswithaclonedppageneofSacharomycescerevisiae.
UnitDefinition: Oneunitistheamountofenzymethatwillgenerate1µmolofphosphateperminutefrominorganicpyrophosphateunderstandardreactionconditions(a10minutereactionat25°Cin100mMTris-HCL,[pH7.2],2mMMgCl2and2mMPPiinareactionvolumeof0.5ml).
StorageConditions: 20mMTris-HCl 100mMKCl 1mMDithiothreitol 0.1mMEDTA 50%Glycerol pH8.0@25°C
RecommendedStorageCondition:-20°C
References: 1.Cooperman,B.S.,Themechanismofactionofyeastinorganicpyrophosphatase,Meth.Enzymol.,87,526-548,1982. 2.Cunningham,P.R.andOfengand,J.,UseofinorganicpyrophostasetoimprovetheyieldofinvitrotranscriptionreactionscatalyzedbyT7RNApolymerase,Biotechniques,9,713-714,1990. 3.Tabor,S.,Richardson,C.C.,DNAsequenceanalysiswithamodifiedbacteriophageT7DNApolymerase.Effectofpyrophosphorolysisandmetalions,J.Biol.Chem.,265,8322-8328,1990. 4.Dean,B.F.,etal.,RapidamplificationofplasmidandphageDNAusingphi29DNApolymeraseandmultiply-primedRollingCircleamplification,GenomeRes.,11,1095-1099,2001. 5.Zhou,G.H.,etal.,Quantitativedetectionofsinglenucleotidepolymorphismsforapooledsamplebyabioluminometricassaycoupledwithmodifiedprimerextensionreactions(BAMPER),NucleicAcidsRes.,29,E93,2001. |