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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 |