YeastPoly(A)PolymeraseusesATPasasubstratefortemplate-independentadditionofAMPfromATPtothe3´hydroxylterminiofRNAmolecules.Itcanbeusedtoaddpoly(A)tailstoRNAinthefirststepofcloning.ThereactionrequiresMn2+orMg2+,ATPassubstrate,andanyRNAcontaining3'hydroxylterminiasprimer.Substitutionofcordycepin-5'-triphosphate(3'-dATP)forATPresultsinadditionofasingle3'-dAresiduetotheendsoftheRNA,ausefultechniqueforlabelingRNAatthe3'end.

Incomparativestudies,YeastPoly(A)PolymeraseworksmoreefficientlythanE.colipoly(A)polymeraseforRNAoligonucleotide-labelingandpoly(A)tailing.Shorterincubationtimesarerequiredfortheyeastenzymeanditisfoundtolabelbothlongandshortsubstratesequallywell.YeastPoly(A)PolymeraseisrecommendedoverT4RNALigasefor3'-endlabelingoflongRNAmolecules.

• Additionofapoly(A)tailtoRNAsynthesizedinvitroforincreasingRNAstABIlityandtranslationefficiencypriortotransferringintoeukaryoticcells

• CDNAlibraryconstructionusingaprimercontainingpoly(dT)onits3´end.

• SynthesisofpolyadenylatedRNAfornucleicacidamplification

• YeastPoly(A)Polymerase:5,000U/mlin20mMTris-HCl,300mMNaCl,1mMDTT,1mMEDTA,50%Glycerol,0.1%Triton®X-100,pH7.5@25°C
• 5XReactionBuffer:1500mMTris-HCl,pH7.0,3.0mMMnCl2,0.1mMEDTA,1mMDTT,500μg/mlacetylatedBSA,50%glycerol.

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