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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 # Name Simultaneous digestion and dephosphorylation Heat inactivation is recommended prior to dephosphorylation Column purification is recommended prior to dephosphorylation
FD2064 FastDigest AanI 1-step    
FD1724 FastDigest AasI 1-step    
FD0994 FastDigest AatII     2-step-column purify first
FD0904 FastDigest Acc65I 1-step    
FD1234 FastDigest AdeI   2-step-heat inactivate first  
FD1954 FastDigest AjuI     2-step-column purify first
FD0014 FastDigest AluI 1-step    
FD0024 FastDigest Alw21I     2-step-column purify first
FD0034 FastDigest Alw26I 1-step    
FD0044 FastDigest Alw44I 1-step    
FD1414 FastDigest ApaI     2-step-column purify first
FD0054 FastDigest BamHI 1-step    
FD0724 FastDigest BclI 1-step    
FD0064 FastDigest BcnI 1-step    
FD1253, FD1254 FastDigest BcuI 1-step    
FD1164 FastDigest BfmI 1-step    
FD0074 FastDigest BglI     2-step-column purify first
FD0083, FD0084 FastDigest BglII 1-step    
FD1424 FastDigest Bme1390I     2-step-column purify first
FD1434 FastDigest BoxI 1-step    
FD1014 FastDigest BpiI 1-step    
FD1314 FastDigest BplI     2-step-column purify first
FD1184 FastDigest Bpu10I     2-step-column purify first
FD0094 FastDigest Bpu1102I   2-step-heat inactivate first  
FD1084 FastDigest BseDI 1-step    
FD0874 FastDigest BseGI   2-step-heat inactivate first  
FD1714 FastDigest BseJI     2-step-column purify first
FD1204 FastDigest BseLI     2-step-column purify first
FD1264 FastDigest BseMI     2-step-column purify first
FD1404 FastDigest H49BseMII     2-step-column purify first
FD0884 FastDigest BseNI   2-step-heat inactivate first  
FD1444 FastDigest BseSI     2-step-column purify first
FD1454 FastDigest BseXI   2-step-heat inactivate first  
FD0924 FastDigest Bsh1236I 1-step    
FD0894 FastDigest Bsh1285I   2-step-heat inactivate first  
FD1004 FastDigest BshNI   2-step-heat inactivate first  
FD1464 FastDigest BshTI     2-step-column purify first
FD0124 FastDigest Bsp119I 1-step    
FD0134 FastDigest Bsp120I 1-step    
FD0933, FD0934 FastDigest Bsp1407I 1-step    
FD0784 FastDigest Bsp143I 1-step    
FD2154 FastDigest RruI     2-step-column purify first
FD1154 FastDigest BspLI 1-step    
FD2044 FastDigest BspOI     2-step-column purify first
FD0834 FastDigest H81BspTI     2-step-column purify first
FD0704 FastDigest Bst1107I     2-step-column purify first
FD1024 FastDigest BstXI     2-step-column purify first
FD0143, FD0144 FastDigest Bsu15I 1-step    
FD0154 FastDigest BsuRI   2-step-heat inactivate first  
FD1744 FastDigest BveI 1-step    
FD1394 FastDigest CaiI     2-step-column purify first
FD0184 FastDigest Cfr10I     2-step-column purify first
FD0194 FastDigest Cfr13I 1-step    
FD0744 FastDigest CpoI 1-step    
FD1904 FastDigest H107CseI     2-step-column purify first
FD0214 FastDigest Csp6I 1-step    
FD1703, FD1704 FastDigest DpnI   2-step-heat inactivate first  
FD0224 FastDigest DraI   2-step-heat inactivate first  
FD0234 FastDigest Eam1104I 1-step    
FD0244 FastDigest Eam1105I 1-step    
FD0254 FastDigest Ecl136II 1-step    
FD0404 FastDigest Eco105I   2-step-heat inactivate first  
FD0414 FastDigest Eco130I 1-step    
FD0424 FastDigest Eco147I 1-step    
FD0293, FD0294 FastDigest Eco31I 1-step    
FD0303, FD0304 FastDigest Eco32I 1-step    
FD0314 FastDigest Eco47I   2-step-heat inactivate first  
FD0324 FastDigest Eco47III   2-step-heat inactivate first  
FD0334 FastDigestEco52I 1-step    
FD0344 FastDigest Eco57I     2-step-column purify first
FD0364 FastDigest Eco72I 1-step    
FD0374 FastDigest Eco81I     2-step-column purify first
FD0384 FastDigest Eco88I   2-step-heat inactivate first  
FD0394 FastDigest Eco91I   2-step-heat inactivate first  
FD0264 FastDigest EcoO109I   2-step-heat inactivate first  
FD0274, FD0275 FastDigest EcoRI   2-step-heat inactivate first  
FD0443, FD0444 FastDigest EheI 1-step    
FD0454 FastDigest Esp3I     2-step-column purify first
FD1814 FastDigest FaqI 1-step    
FD1664 FastDigest FspAI 1-step    
FD1764 FastDigest FspBI 1-step    
FD0464 FastDigest GsuI     2-step-column purify first
FD1854 FastDigest HhaI 1-step    
FD0474 FastDigest Hin1I     2-step-column purify first
FD1834 FastDigest Hin1II     2-step-column purify first
FD0484 FastDigest Hin6I   2-step-heat inactivate first  
FD0494 FastDigest HincII 1-step    
FD0504, FD5005 FastDigest HindIII     2-step-column purify first
FD0804 FastDigest HinfI 1-step    
FD0514 FastDigest HpaII 1-step    
FD1574 FastDigest Hpy8I 1-step    
FD1734 FastDigest HpyF10VI     2-step-column purify first
FD1884 FastDigest HpyF3I 1-step    
FD0534 FastDigest Kpn2I   2-step-heat inactivate first  
FD0524 FastDigest KpnI     2-step-column purify first
FD1034 FastDigest KspAI   2-step-heat inactivate first  
FD1934 FastDigest LguI 1-step    
FD2074 FastDigest Lsp1109I 1-step    
FD2124 FastDigest BmsI 1-step    
FD2084 FastDigest MauBI 1-step    
FD1274 FastDigest MbiI 1-step    
FD0814 FastDigest MboI 1-step    
FD0824 FastDigest MboII 1-step    
FD1214 FastDigest MlsI 1-step    
FD0564 FastDigest MluI   2-step-heat inactivate first  
FD1074 FastDigest MnlI 1-step    
FD0734 FastDigest Mph1103I     2-step-column purify first
FD2024 FastDigest MreI 1-step    
FD0544 FastDigest MspI 1-step    
FD1344 FastDigest MssI 1-step    
FD0753, FD0754 FastDigest MunI 1-step    
FD0964 FastDigest Mva1269I     2-step-column purify first
FD0554 FastDigest MvaI     2-step-column purify first
FD0573, FD0574, FD0575 FastDigest NcoI 1-step    
FD0583, FD0584, FD0585 FastDigest NdeI 1-step    
FD0973, FD0974 FastDigest NheI 1-step    
FD1514 FastDigest NmuCI 1-step    
FD0593, FD0594, FD0595, FD0596 FastDigest NotI 1-step    
FD1224 FastDigest NsbI 1-step    
FD1634 FastDigest OliI 1-step    
FD2204 FastDigest PacI     2-step-column purify first
FD0604 FastDigest PaeI     2-step-column purify first
FD1284 FastDigest PagI 1-step    
FD2134 FastDigest PteI   2-step-heat inactivate first  
FD1524 FastDigest PdiI 1-step    
FD1534 FastDigest PdmI 1-step    
FD1784 FastDigest PfeI 1-step    
FD0854 FastDigest Pfl23II 1-step    
FD1754 FastDigest PfoI   2-step-heat inactivate first  
FD1974 FastDigest BsaAI (Ppu21I)   2-step-heat inactivate first  
FD0944 FastDigest AclI (Psp1406I) 1-step    
FD0764 FastDigest PpuMI (Psp5II)     2-step-column purify first
FD0614, FD06145 FastDigest PstI     2-step-column purify first
FD1554 FastDigest PsuI     2-step-column purify first
FD1334 FastDigest PsyI     2-step-column purify first
FD0624 FastDigest PvuI 1-step    
FD0634 FastDigest PvuII 1-step    
FD1124 FastDigest RsaI 1-step    
FD2004 FastDigest RseI 1-step    
FD1133, FD1134 FastDigest SacI     2-step-column purify first
FD0644 FastDigest SalI 1-step    
FD1644 FastDigest SatI     2-step-column purify first
FD0434 FastDigest ScaI 1-step    
FD1374 FastDigest SchI     2-step-column purify first
FD1194 FastDigest SdaI     2-step-column purify first
FD0654 FastDigest SduI     2-step-column purify first
FD2094 FastDigest SfaAI 1-step    
FD1824 FastDigest SfiI     2-step-column purify first
FD1894 FastDigest SgsI 1-step    
FD0663, FD0664 FastDigest SmaI 1-step    
FD1244 FastDigest SmiI 1-step    
FD1794 FastDigest SsiI 1-step    
FD0774 FastDigest SspI 1-step    
FD1364 FastDigest TaaI     2-step-column purify first
FD1144 FastDigest TaiI     2-step-column purify first
FD0674 FastDigest TaqI     2-step-column purify first
FD1354 FastDigest TasI     2-step-column purify first
FD1294 FastDigest TatI     2-step-column purify first
FD165 FastDigest TauI     2-step-column purify first
FD0984 FastDigest Tru1I     2-step-column purify first
FD2104 FastDigest TscAI     2-step-column purify first
FD0714 FastDigest Van91I     2-step-column purify first
FD0914 FastDigest VspI 1-step    
FD1304 FastDigest XagI   2-step-heat inactivate first  
FD1383, FD1384 FastDigest XapI   2-step-heat inactivate first  
FD0684, FD0685 FastDigest XbaI 1-step    
FD1474 FastDigest XceI     2-step-column purify first
FD0694, FD0695 FastDigest XhoI 1-step    
FD1564 FastDigest XmaJI 1-step    
FD1484 FastDigest XmiI     2-step-column purify first
FD2144 FastDigest FokI 1-step    
FD2164 FastDigest KflI 1-step    
FD2224 FastDigest PspFI     2-step-column purify first
FD2114 FastDigest CsiI     2-step-column purify first
FD2174 FastDigest SaqAI     2-step-column purify first
FD2184 FastDigest BfoI 1-step