TheExonucleaseI(ExoI)degradessingle-strandedDNAina3'=>5'direction,releasingdeoxyribonucleoside5'-monophosphatesinastepwisemannerandleaving5'-terminaldinucleotidesintact.ItdoesnotcleaveDNAstrandswithterminal3'-OHgroupsblockedbyphosphoryloracetylgroups.||Source:|E.colicellswithaclonedE.colisbcBgene.||Concentration:~20u/ul||UnitDefinition:|OneunitofE9007-85Ccatalyzesthereleaseof10nano-molesofacidsolublenucleotidesin30minat37ºC.||ActivityAssay:|67mMglycine-KOHpH9.5,6.7mMMgCl2,1mMDTT,0.17mg/mlsinglestranded[3H]-DNA.||Form:|Suppliedasaliquidin20mMTris-HCl,pH7.5,0.1mMEDTA,1mMDTTand50%glycerol.||Suppliedwith:|E9007-85C1:10XReactionBuffer:Suppliedasaliquidin670mMglycine-KOH,pH9.5at25°C,67mMMgCl2,10mMDTT.||Applications:|SuitableforuseinthepurificationofPCRproducts,removalofsingle-strandedDNAcontaininga3'-hydroxylterminusfromnucleicacidmixturesandassayforthepresenceofsingle-strandedDNAwitha3'-hydroxylterminus.Otherapplicationsnottested.||ProtocolforEliminationofOligonucleotidesanddNTPsfromPCTmixtures:|1.Removea5ulaliquotfromareactionmixture.|2.Add10uofE9007-85Cand1uofShrimpAlkalinePhosphatase.|3.Mixandincubateat37ºCfor15min.|4.Inactivatetheenzymesbyheatingat85ºCfor15min.|*Note:Afterinactivation,thealiquotcanbeuseddirectlyforDNAsequencing.TheExonucleaseIisnotsuitableforremoving3’-overhangsofdsDNA.||StorageandStABIlity:|Maybestoredat4°Cforshort-termonly.Aliquottoavoidrepeatedfreezingandthawing.Storeat-20°C.Aliquotsarestablefor6monthsafterreceipt.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.

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