Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
Reaction Conditions for FastDigest Restriction Enzymes |
The reaction conditions table provides detailed information about the optimal reaction conditions for each FastDigest restriction enzyme, including recommended buffers, incubation times, and temperatures. This comprehensive overview helps molecular biologists in achieving efficient and precise DNA digestion, minimizing the need for extensive optimization.
Thermo Scientific FastDigest enzyme | Reaction temperature | Digestion time with 1 µL of FastDigest enzyme, min | bp from end of DNA required for complete digestion | Thermal inactivation | Incubation time (in hours) without star activity | |||
---|---|---|---|---|---|---|---|---|
Lambda, 1 µg/20 µL | Plasmid DNA, 1 µg/20 µL | PCR product, ~0.2 µg/30 µL | Genomic DNA, 1 µg/10 µL | |||||
FastDigest AatII | 37°C | 15 (pUC19 DNA/SmaI) | 20 | 15 | 15 | 5 | 80°C, 5 min | 16 |
FastDigest AanI | 37°C | 5 | 5 | 5 | 5 | 5 | 65°C, 5 min | 16 |
FastDigest AasI | 37°C | 5 | 5 | 5 | 5 | 1 | 80°C, 10 min | 1 |
FastDigest Acc65I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest AdeI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 1 |
FastDigest AjuI | 37°C | 5 (phiX174 DNA) | 5 | 10 | 5 | nd | 65°C, 5 min | 16 |
FastDigest AluI | 37°C | 15 | 15 | 15 | 15 | 4 | 65°C, 5 min | 16 |
FastDigest Alw21I | 37°C | 5 | 5 | 5 | 5 | 1 | 80°C, 20 min | 16 |
FastDigest Alw26I | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest Alw44I | 37°C | 5 | 60 | 10 | 5 | 4 | 80°C, 5 min | 16 |
FastDigest ApaI | 37°C | 5 | 5 | 20 | 10 | 2 | 65°C, 5 min | 16 |
FastDigest BamHI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 1 |
FastDigest BclI | 37°C | 5 | 5 | 15 | 5 | 3 | 80°C, 20 min | 16 |
FastDigest BcnI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 20 min | 16 |
FastDigest BcuI | 37°C | no sites (pUC19-SpeI DNA/Psp1406I, 5 min) | 5 | 5 | 5 | 1 | No (chloroform extraction) | 16 |
FastDigest BfmI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 20 min | 16 |
FastDigest BfoI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 10 min | 16 |
FastDigest BglI | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 2 |
FastDigest BglII | 37°C | 5 | 20 | 30 | 20 | 3 | No (chloroform extraction) | 16 |
FastDigest Bme1390I | 37°C | 5 | 5 | 5 | 5 | 4 | No (chloroform extraction) | 16 |
FastDigest BmsI | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest BoxI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 20 min | 16 |
FastDigest BpiI | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 10 min | 16 |
FastDigest BplI | 37°C | 15 (lambda DNA/XhoI) | 20 | 30 | 30 | nd | 65°C, 5 min | 16 |
FastDigest Bpu10I | 37°C | 15 (M13mp18 DNA) | 15 | 30 | 15 | 3 | 80°C, 5 min | 1 |
FastDigest Bpu1102I | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest BseDI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 5 min | 16 |
FastDigest BseGI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest BseJI | 65°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 1 |
FastDigest BseLI | 37°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 16 |
FastDigest BseMI | 37°C | 15 | 15 | 15 | 15 | 3 | 80°C, 5 min | 16 |
FastDigest BseMII | 55°C | 15 | 30 | 15 | 15 | 2 | 80°C, 5 min | 16 |
FastDigest BseNI | 65°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest BseSI | 37°C | 5 | 5 | 5 | 30 | 3 | 80°C, 15 min | 16 |
FastDigest BseXI | 65°C | 15 (pBR322 DNA) | 15 | 15 | 15 | 3 | 80°C, 15 min | 16 |
FastDigest Bsh1236I | 37°C | 5 | 5 | 5 | 5 | 1 | 80°C, 10 min | 16 |
FastDigest Bsh1285I | 37°C | 15 | 15 | 15 | 15 | 3 | 80°C, 15 min | 1 |
FastDigest BshNI | 37°C | 5 | 60 | 5 | 15 | 2 | 65°C, 10 min | 16 |
FastDigest BshTI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 5 min | 16 |
FastDigest Bsp119I | 37°C | 5 | 5 | 5 | 5 | 1 | 80°C, 5 min | 16 |
FastDigest Bsp120I | 37°C | 5 | 5 | 15 | 5 | 3 | 80°C, 10 min | 16 |
FastDigest Bsp1407I | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 10 min | 16 |
FastDigest Bsp143I | 37°C | 5 | 5 | 10 | 5 | 4 | 65°C, 20 min | 16 |
FastDigest BspLI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 20 min | 16 |
FastDigest BspOI | 37°C | 5 | 5 | 15 | 5 | 3 | 80°C, 5 min | 1 |
FastDigest BspTI | 37°C | 5 | 5 | 5 | 5 | 4 | No (chloroform extraction) | 16 |
FastDigest Bst1107I | 37°C | 5 | 5 | 5 | 10 | 3 | No (chloroform extraction) | 6 |
FastDigest BstXI | 37°C | 5 | 5 | 5 | 5 | 4 | 80°C, 5 min | 1 |
FastDigest Bsu15I | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 15 min | 16 |
FastDigest BsuRI | 37°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest BveI | 37°C | 15 | 15 | 60 | 20 | 5 | 65°C, 5 min | 16 |
FastDigest CaiI | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 10 min | 16 |
FastDigest Cfr10I | 37°C | 5 | 5 | 10 | 5 | 2 | No (chloroform extraction) | 2 |
FastDigest Cfr13I | 37°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 16 |
FastDigest CpoI | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest CseI | 37°C | 15 (pBR322 DNA) | 15 | 30 | 15 | 2 | 80°C, 5 min | 16 |
FastDigest CsiI | 37°C | 5 | 10 | 5 | 15 | 4 | 65°C, 5 min | 16 |
FastDigest Csp6I | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 10 min | 16 |
FastDigest DpnI | 37°C | 5 (pBR322 DNA) | 5 | not applicable | 10 | 1 | 80°C, 5 min | 16 |
FastDigest DraI | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 5 min | 16 |
FastDigest Eam1104I | 37°C | 5 (phiX174 DNA) | 5 | 5 | 5 | 3 | 80°C, 5 min | 6 |
FastDigest Eam1105I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Ecl136II | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 6 |
FastDigest Eco105I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Eco130I | 37°C | 5 | 5 | 20 | 20 | 3 | 65°C, 5 min | 16 |
FastDigest Eco147I | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 10 min | 16 |
FastDigest Eco31I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Eco32I | 37°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 16 |
FastDigest Eco47I | 37°C | 5 | 5 | 5 | 5 | 1 | 80°C, 20 min | 16 |
FastDigest Eco47III | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 5 min | 16 |
FastDigest Eco52I | 37°C | 5 | 20 | 20 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Eco57I | 37°C | 15 | 30 | 15 | 15 | 2 | 65°C, 5 min | 16 |
FastDigest Eco72I | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 10 min | 1 |
FastDigest Eco81I | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 10 min | 16 |
FastDigest Eco88I | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest Eco91I | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 10 min | 2 |
FastDigest EcoO109I | 37°C | 5 | 10 | 15 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest EcoRI | 37°C | 5 | 5 | 20 | 5 | 2 | 80°C, 5min | 0.5 |
FastDigest EheI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 6 |
FastDigest Esp3I | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 10 min | 6 |
FastDigest FaqI | 37°C | 15 | 30 | 15 | 15 | 3 | 65°C, 5 min | 16 |
FastDigest FokI | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 1 |
FastDigest FspAI | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 5 min | 16 |
FastDigest FspBI | 37°C | 5 | 15 | 5 | 20 | 3 | 80°C, 5 min | 16 |
FastDigest GsuI | 30°C | 15 | 30 | 15 | 15 | 2 | 65°C, 5 min | 16 |
FastDigest HhaI | 37°C | 5 | 5 | 5 | 5 | 1 | No (chloroform extraction) | 16 |
FastDigest Hin1I | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest Hin1II | 37°C | 5 | 10 | 5 | 10 | 4 | 80°C, 5 min | 16 |
FastDigest Hin6I | 37°C | 5 | 10 | 5 | 5 | 2 | 80°C, 10 min | 16 |
FastDigest HincII | 37°C | 5 | 5 | 5 | 10 | 1 | 65°C, 5 min | 16 |
FastDigest HindIII | 37°C | 5 | 5 | 20 | 10 | 3 | 80°C, 10 min | 16 |
FastDigest HinfI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 20 min | 16 |
FastDigest HpaII | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Hpy8I | 37°C | 5 | 5 | 5 | 10 | 2 | 80°C, 5 min | 16 |
FastDigest HpyF10VI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest HpyF3I | 37°C | 5 | 5 | 5 | 10 | 3 | 65°C, 5 min | 16 |
FastDigest KflI | 37°C | 5 (pJET1-SanDI DNA/BamHI) | 5 | 5 | 10 | 5 | No (chloroform extraction) | 16 |
FastDigest Kpn2I | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest KpnI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 5 min | 16 |
FastDigest KspAI | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 20 min | 0.5 |
FastDigest LguI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest Lsp1109I | 37°C | 5 (pBR322 DNA) | 5 | 5 | 10 | 2 | 65°C, 5 min | 1 |
FastDigest MauBI | 37°C | no sites (pJET1-MauBI DNA/BamHI, 5 min) | 5 | 10 | 5 | 5 | 65°C, 5 min | 16 |
FastDigest MbiI | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest MboI | 37°C | 5 | 5 | 5 | 10 | 1 | 65°C, 15 min | 16 |
FastDigest MboII | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest MlsI | 37°C | 5 | 5 | 30 | 20 | 3 | 80°C, 20 min | 16 |
FastDigest MluI | 37°C | 5 | 5 | 5 | 15 | 3 | 80°C, 5 min | 16 |
FastDigest MnlI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest Mph1103I | 37°C | 5 | 5 | 5 | 10 | 3 | 65°C, 15 min | 6 |
FastDigest MreI | 37°C | no sites (pJET1-MreI DNA/Eam1105I, 5 min) | 5 | 5 | 5 | 3 | 80°C, 5 min | 16 |
FastDigest MspI | 37°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest MssI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 10 min | 16 |
FastDigest MunI | 37°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest Mva1269I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest MvaI | 37°C | 5 | 5 | 5 | 10 | 4 | No (chloroform extraction) | 1 |
FastDigest NcoI | 37°C | 5 | 10 | 10 | 5 | 3 | 65°C, 15 min | 16 |
FastDigest NdeI | 37°C | 5 | 5 | 60 | 30 | 3 | 65°C, 5 min | 6 |
FastDigest NheI | 37°C | 5 | 15 | 5 | 5 | 5 | 65°C, 5 min | 6 |
FastDigest NmuCI | 37°C | 5 | 5 | 5 | 10 | 2 | 65°C, 5 min | 16 |
FastDigest NotI | 37°C | no sites (pTZ19RJL2 DNA/BseLI, 5 min) | 30 | 5 | 10 | 2 | 80°C, 5 min | 16 |
FastDigest NsbI | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 15 min | 16 |
FastDigest OliI | 37°C | 5 (phiX174 DNA) | 5 | 10 | 5 | 3 | 65°C, 5 min | 6 |
FastDigest PacI | 37°C | no sites (pJET1-PacI DNA/SdaI, 5 min) | 5 | 5 | 5 | 2 | 65°C, 10 min | 16 |
FastDigest PaeI | 37°C | 5 | 5 | 5 | 5 | 5 | 65°C, 5 min | 16 |
FastDigest PagI | 37°C | 5 (phiX174 DNA) | 10 | 5 | 10 | 3 | 80°C, 5 min | 0.5 |
FastDigest PdiI | 37°C | 5 (pBR322 DNA/NdeI) | 5 | 5 | 5 | 4 | 65°C, 15 min | 16 |
FastDigest PdmI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest PfeI | 37°C | 5 | 5 | 10 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest Pfl23II | 37°C | 15 | 15 | 15 | 15 | 3 | 65°C, 5 min | 16 |
FastDigest PfoI | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Ppu21I | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest PsyI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 4 |
FastDigest Psp1406I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest Psp5II | 37°C | 5 | 5 | 5 | 20 | 2 | 80°C, 5 min | 6 |
FastDigest PspFI | 37°C | 5 | 5 | 5 | 5 | 4 | 80°C, 5 min | 16 |
FastDigest PstI | 37°C | 5 | 5 | 30 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest PsuI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 5 min | 16 |
FastDigest PteI | 37°C | 5 | 5 | 5 | 5 | 3 | 80°C, 5 min | 16 |
FastDigest PvuI | 37°C | 5 | 15 | 5 | 5 | 4 | 80°C, 5 min | 16 |
FastDigest PvuII | 37°C | 5 | 5 | 5 | 5 | 1 | No (chloroform extraction) | 0.5 |
FastDigest RruI | 37°C | 5 | 5 | 5 | 5 | 5 | No (chloroform extraction) | 4 |
FastDigest RsaI | 37°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest RseI | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 20 min | 16 |
FastDigest SacI | 37°C | 5 | 15 | 30 | 10 | 1 | 65°C, 5 min | 16 |
FastDigest SalI | 37°C | 5 | 5 | 60 | 5 | 3 | 65°C, 10 min | 16 |
FastDigest SaqAI | 37°C | 5 (pBR322 DNA) | 5 | 5 | 5 | 4 | 65°C, 5 min | 1 |
FastDigest SatI | 37°C | 5 | 5 | 10 | 5 | 3 | 65°C, 5 min | 16 |
FastDigest ScaI | 37°C | 5 | 5 | 5 | 5 | 4 | 65°C, 10 min | 16 |
FastDigest SchI | 37°C | 5 | 5 | 30 | 5 | 2 | 80°C, 5 min | 1 |
FastDigest SdaI | 37°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 1 |
FastDigest SduI | 37°C | 5 | 5 | 5 | 5 | 2 | 80°C, 15 min | 1 |
FastDigest SfaAI | 37°C | no sites (pJET1-AsiSI DNA/SdaI, 5 min) | 5 | 60 | 5 | 5 | 80°C, 5 min | 16 |
FastDigest SfiI | 50°C | no sites (pUC19-SfiI DNA, 15 min) | 15 | 15 | 15 | 5 | No (chloroform extraction) | 16 |
FastDigest SgsI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 20 min | 16 |
FastDigest SmaI | 37°C | 5 | 5 | 5 | 5 | 1 | 65°C, 5 min | 16 |
FastDigest SmiI | 37°C | no sites (M13mp18 DNA/MvaI, 5 min) | 20 | 30 | 30 | 2 | 65°C, 15 min | 1 |
FastDigest SsiI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 4 |
FastDigest SspI | 37°C | 5 | 5 | 5 | 30 | 2 | 65°C, 5 min | 1 |
FastDigest TaaI | 65°C | 5 | 5 | 5 | 5 | 3 | No (chloroform extraction) | 16 |
FastDigest TaiI | 65°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 16 |
FastDigest TaqI | 65°C | 5 | 5 | 5 | 5 | 4 | No (chloroform extraction) | 16 |
FastDigest TasI | 65°C | 5 (pBR322 DNA) | 5 | 5 | 5 | 3 | No (chloroform extraction) | 6 |
FastDigest TatI | 65°C | 15 | 15 | 15 | 20 | 5 | No (chloroform extraction) | 1 |
FastDigest TauI | 55°C | 15 | 60 | 15 | 30 | 3 | No (chloroform extraction) | 16 |
FastDigest Tru1I | 65°C | 5 | 5 | 5 | 10 | 3 | No (chloroform extraction) | 2 |
FastDigest TscAI | 65°C | 5 | 5 | 5 | 5 | 2 | No (chloroform extraction) | 6 |
FastDigest Van91I | 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 10 min | 6 |
FastDigest VspI | 37°C | 5 | 5 | 5 | 30 | 2 | 65°C, 5 min | 16 |
FastDigest XagI | 37°C | 5 | 5 | 5 | 20 | 2 | 65°C, 5 min | 16 |
FastDigest XapI | 37°C | 5 | 5 | 5 | 10 | 4 | 80°C, 5 min | 16 |
FastDigest XbaI | 37°C | 5 | 5 | 5 | 10 | 2 | 65°C, 20 min | 16 |
FastDigest XceI | 37°C | 5 | 5 | 5 | 5 | 2 | 65°C, 5 min | 16 |
FastDigest XhoI | 37°C | 5 | 5 | 5 | 10 | 2 | 80°C, 5 min | 16 |
FastDigest XmaJI | 37°C | 5 | 10 | 5 | 10 | 2 | No (chloroform extraction) | 16 |
FastDigest XmiI | 37°C | 5 | 5 | 60 | 5 | 2 | 65°C, 5 min | 16 |
For Research Use Only. Not for use in diagnostic procedures.