For years we have known that FastAP Alkaline Phosphatase is fully active in the FastDigest Buffers, both green and colorless versions. However, recent thorough examination of the enzymes revealed that dephosphorylation efficiency is highly dependent upon the nature of the restriction enzyme used in digestion.

With some enzymes, restriction digestion and dephosphorylation reactions can be performed simultaneously. Others may require heat inactivation or column purification following restriction enzyme digestion and prior to the addition of FastAP Alkaline Phosphatase.

As a result, all FastDigest Restriction Enzymes have been tested to determine the optimal protocol for complete dephosphorylation when using FastAP Alkaline Phosphatase. The recommendations are outlined in the table below.

Catalog #NameSimultaneous digestion and dephosphorylationHeat inactivation is recommended prior to dephosphorylationColumn purification is recommended prior to dephosphorylation
FD2064FastDigest AanI1-step  
FD1724FastDigest AasI1-step  
FD0994FastDigest AatII  2-step-column purify first
FD0904FastDigest Acc65I1-step  
FD1234FastDigest AdeI 2-step-heat inactivate first 
FD1954FastDigest AjuI  2-step-column purify first
FD0014FastDigest AluI1-step  
FD0024FastDigest Alw21I  2-step-column purify first
FD0034FastDigest Alw26I1-step  
FD0044FastDigest Alw44I1-step  
FD1414FastDigest ApaI  2-step-column purify first
FD0054FastDigest BamHI1-step  
FD0724FastDigest BclI1-step  
FD0064FastDigest BcnI1-step  
FD1253, FD1254FastDigest BcuI1-step  
FD1164FastDigest BfmI1-step  
FD0074FastDigest BglI  2-step-column purify first
FD0083, FD0084FastDigest BglII1-step  
FD1424FastDigest Bme1390I  2-step-column purify first
FD1434FastDigest BoxI1-step  
FD1014FastDigest BpiI1-step  
FD1314FastDigest BplI  2-step-column purify first
FD1184FastDigest Bpu10I  2-step-column purify first
FD0094FastDigest Bpu1102I 2-step-heat inactivate first 
FD1084FastDigest BseDI1-step  
FD0874FastDigest BseGI 2-step-heat inactivate first 
FD1714FastDigest BseJI  2-step-column purify first
FD1204FastDigest BseLI  2-step-column purify first
FD1264FastDigest BseMI  2-step-column purify first
FD1404FastDigest H49BseMII  2-step-column purify first
FD0884FastDigest BseNI 2-step-heat inactivate first 
FD1444FastDigest BseSI  2-step-column purify first
FD1454FastDigest BseXI 2-step-heat inactivate first 
FD0924FastDigest Bsh1236I1-step  
FD0894FastDigest Bsh1285I 2-step-heat inactivate first 
FD1004FastDigest BshNI 2-step-heat inactivate first 
FD1464FastDigest BshTI  2-step-column purify first
FD0124FastDigest Bsp119I1-step  
FD0134FastDigest Bsp120I1-step  
FD0933, FD0934FastDigest Bsp1407I1-step  
FD0784FastDigest Bsp143I1-step  
FD2154FastDigest RruI  2-step-column purify first
FD1154FastDigest BspLI1-step  
FD2044FastDigest BspOI  2-step-column purify first
FD0834FastDigest H81BspTI  2-step-column purify first
FD0704FastDigest Bst1107I  2-step-column purify first
FD1024FastDigest BstXI  2-step-column purify first
FD0143, FD0144FastDigest Bsu15I1-step  
FD0154FastDigest BsuRI 2-step-heat inactivate first 
FD1744FastDigest BveI1-step  
FD1394FastDigest CaiI  2-step-column purify first
FD0184FastDigest Cfr10I  2-step-column purify first
FD0194FastDigest Cfr13I1-step  
FD0744FastDigest CpoI1-step  
FD1904FastDigest H107CseI  2-step-column purify first
FD0214FastDigest Csp6I1-step  
FD1703, FD1704FastDigest DpnI 2-step-heat inactivate first 
FD0224FastDigest DraI 2-step-heat inactivate first 
FD0234FastDigest Eam1104I1-step  
FD0244FastDigest Eam1105I1-step  
FD0254FastDigest Ecl136II1-step  
FD0404FastDigest Eco105I 2-step-heat inactivate first 
FD0414FastDigest Eco130I1-step  
FD0424FastDigest Eco147I1-step  
FD0293, FD0294FastDigest Eco31I1-step  
FD0303, FD0304FastDigest Eco32I1-step  
FD0314FastDigest Eco47I 2-step-heat inactivate first 
FD0324FastDigest Eco47III 2-step-heat inactivate first 
FD0344FastDigest Eco57I  2-step-column purify first
FD0364FastDigest Eco72I1-step  
FD0374FastDigest Eco81I  2-step-column purify first
FD0384FastDigest Eco88I 2-step-heat inactivate first 
FD0394FastDigest Eco91I 2-step-heat inactivate first 
FD0264FastDigest EcoO109I 2-step-heat inactivate first 
FD0274, FD0275FastDigest EcoRI 2-step-heat inactivate first 
FD0443, FD0444FastDigest EheI1-step  
FD0454FastDigest Esp3I  2-step-column purify first
FD1814FastDigest FaqI1-step  
FD1664FastDigest FspAI1-step  
FD1764FastDigest FspBI1-step  
FD0464FastDigest GsuI  2-step-column purify first
FD1854FastDigest HhaI1-step  
FD0474FastDigest Hin1I  2-step-column purify first
FD1834FastDigest Hin1II  2-step-column purify first
FD0484FastDigest Hin6I 2-step-heat inactivate first 
FD0494FastDigest HincII1-step  
FD0504, FD5005FastDigest HindIII  2-step-column purify first
FD0804FastDigest HinfI1-step  
FD0514FastDigest HpaII1-step  
FD1574FastDigest Hpy8I1-step  
FD1734FastDigest HpyF10VI  2-step-column purify first
FD1884FastDigest HpyF3I1-step  
FD0534FastDigest Kpn2I 2-step-heat inactivate first 
FD0524FastDigest KpnI  2-step-column purify first
FD1034FastDigest KspAI 2-step-heat inactivate first 
FD1934FastDigest LguI1-step  
FD2074FastDigest Lsp1109I1-step  
FD2124FastDigest BmsI1-step  
FD2084FastDigest MauBI1-step  
FD1274FastDigest MbiI1-step  
FD0814FastDigest MboI1-step  
FD0824FastDigest MboII1-step  
FD1214FastDigest MlsI1-step  
FD0564FastDigest MluI 2-step-heat inactivate first 
FD1074FastDigest MnlI1-step  
FD0734FastDigest Mph1103I  2-step-column purify first
FD2024FastDigest MreI1-step  
FD0544FastDigest MspI1-step  
FD1344FastDigest MssI1-step  
FD0753, FD0754FastDigest MunI1-step  
FD0964FastDigest Mva1269I  2-step-column purify first
FD0554FastDigest MvaI  2-step-column purify first
FD0573, FD0574, FD0575FastDigest NcoI1-step  
FD0583, FD0584, FD0585FastDigest NdeI1-step  
FD0973, FD0974FastDigest NheI1-step  
FD1514FastDigest NmuCI1-step  
FD0593, FD0594, FD0595, FD0596FastDigest NotI1-step  
FD1224FastDigest NsbI1-step  
FD1634FastDigest OliI1-step  
FD2204FastDigest PacI  2-step-column purify first
FD0604FastDigest PaeI  2-step-column purify first
FD1284FastDigest PagI1-step  
FD2134FastDigest PteI 2-step-heat inactivate first 
FD1524FastDigest PdiI1-step  
FD1534FastDigest PdmI1-step  
FD1784FastDigest PfeI1-step  
FD0854FastDigest Pfl23II1-step  
FD1754FastDigest PfoI 2-step-heat inactivate first 
FD1974FastDigest BsaAI (Ppu21I) 2-step-heat inactivate first 
FD0944FastDigest AclI (Psp1406I)1-step  
FD0764FastDigest PpuMI (Psp5II)  2-step-column purify first
FD0614, FD06145FastDigest PstI  2-step-column purify first
FD1554FastDigest PsuI  2-step-column purify first
FD1334FastDigest PsyI  2-step-column purify first
FD0624FastDigest PvuI1-step  
FD0634FastDigest PvuII1-step  
FD1124FastDigest RsaI1-step  
FD2004FastDigest RseI1-step  
FD1133, FD1134FastDigest SacI  2-step-column purify first
FD0644FastDigest SalI1-step  
FD1644FastDigest SatI  2-step-column purify first
FD0434FastDigest ScaI1-step  
FD1374FastDigest SchI  2-step-column purify first
FD1194FastDigest SdaI  2-step-column purify first
FD0654FastDigest SduI  2-step-column purify first
FD2094FastDigest SfaAI1-step  
FD1824FastDigest SfiI  2-step-column purify first
FD1894FastDigest SgsI1-step  
FD0663, FD0664FastDigest SmaI1-step  
FD1244FastDigest SmiI1-step  
FD1794FastDigest SsiI1-step  
FD0774FastDigest SspI1-step  
FD1364FastDigest TaaI  2-step-column purify first
FD1144FastDigest TaiI  2-step-column purify first
FD0674FastDigest TaqI  2-step-column purify first
FD1354FastDigest TasI  2-step-column purify first
FD1294FastDigest TatI  2-step-column purify first
FD165FastDigest TauI  2-step-column purify first
FD0984FastDigest Tru1I  2-step-column purify first
FD2104FastDigest TscAI  2-step-column purify first
FD0714FastDigest Van91I  2-step-column purify first
FD0914FastDigest VspI1-step  
FD1304FastDigest XagI 2-step-heat inactivate first 
FD1383, FD1384FastDigest XapI 2-step-heat inactivate first 
FD0684, FD0685FastDigest XbaI1-step  
FD1474FastDigest XceI  2-step-column purify first
FD0694, FD0695FastDigest XhoI1-step  
FD1564FastDigest XmaJI1-step  
FD1484FastDigest XmiI  2-step-column purify first
FD2144FastDigest FokI1-step  
FD2164FastDigest KflI1-step  
FD2224FastDigest PspFI  2-step-column purify first
FD2114FastDigest CsiI  2-step-column purify first
FD2174FastDigest SaqAI  2-step-column purify first
FD2184FastDigest BfoI1-step  

For Research Use Only. Not for use in diagnostic procedures.