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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, 1µg/20µL Plasmid DNA, 1µg/20µL PCR product, ~0.2µg/30µL Genomic DNA, 1µg/10µL
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 65°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, 5 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

Note

  • (other control substrate DNA):
    • FastDigest AatI assayed using pUC19 DNA
    • FastDigest AjuI assayed using phiX174 DNA
    • FastDigest OliI assayed using phiX174 DNA
    • FastDigest SfaAI assayed using pJET1 DNA containing an insert with a SfaAI recognition site
    • FastDigest Lsp1109I assayed using pBR322 DNA
    • FastDigest Bpu10I assayed using M13mp18 DNA
    • FastDigest BseXI assayed using pBR322 DNA
    • FastDigest PagI assayed using phiX174 DNA
    • FastDigest DpnI, which requires the presence of N6-methyladenine within the recognition sequence, assayed using dam+ methylated pBR322 DNA
    • FastDigest Eam1104I assayed using phiX174 DNA
    • FastDigest CseI assayed using pBR322 DNA
    • FastDigest MauBI assayed using pJET1 DNA containing an insert with a MauBI recognition site
    • FastDigest MreI assayed using pJET1 DNA containing an insert with a MreI recognition site
    • FastDigest MseI assayed using pBR322 DNA
    • FastDigest PdiI assayed using pBR322 DNA
    • FastDigest NotI assayed using pTZ19RJL2 DNA
    • FastDigest KflI assayed using pJET1 DNA containing an insert with a SanDI recognition site
    • FastDigest SfiI assayed using pUC19 DNA containing an insert with a SfiI recognition site
    • FastDigest BcuI assayed using pUC19 DNA containing an insert with a SpeI recognition site
    • FastDigest SmiI assayed using M13mp18 DNA
    • FastDigest TasI assayed using pBR322 DNA
  • No (chloroform extraction) – phenol/chloroform extraction and ethanol precipitation of DNA is recommended to inactivate the enzyme.