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Thermo Scientific FastDigest enzymeReaction temperatureDigestion time with 1 µL of FastDigest enzyme, minbp from end of DNA required for complete digestionThermal inactivationIncubation time (in hours) without star activity
Lambda, 1 µg/20 µLPlasmid DNA, 1 µg/20 µLPCR product, ~0.2 µg/30 µLGenomic DNA, 1 µg/10 µL
FastDigest AatII37°C15
(pUC19 DNA/SmaI)
201515580°C, 5 min16
FastDigest AanI37°C5555565°C, 5 min16
FastDigest AasI37°C5555180°C, 10 min1
FastDigest Acc65I37°C5555365°C, 5 min16
FastDigest AdeI37°C5555280°C, 5 min1
FastDigest AjuI37°C5
(phiX174 DNA)
5105nd65°C, 5 min16
FastDigest AluI37°C15151515465°C, 5 min16
FastDigest Alw21I37°C5555180°C, 20 min16
FastDigest Alw26I37°C5555165°C, 5 min16
FastDigest Alw44I37°C560105480°C, 5 min16
FastDigest ApaI37°C552010265°C, 5 min16
FastDigest BamHI37°C5555280°C, 5 min1
FastDigest BclI37°C55155380°C, 20 min16
FastDigest BcnI37°C5555380°C, 20 min16
FastDigest BcuI37°Cno sites
(pUC19-SpeI DNA/Psp1406I, 5 min)
5551No (chloroform extraction)16
FastDigest BfmI37°C5555265°C, 20 min16
FastDigest BfoI37°C5555265°C, 10 min16
FastDigest BglI37°C5555365°C, 5 min2
FastDigest BglII37°C52030203No (chloroform extraction)16
FastDigest Bme1390I37°C55554No (chloroform extraction)16
FastDigest BmsI37°C5555165°C, 5 min16
FastDigest BoxI37°C5555380°C, 20 min16
FastDigest BpiI37°C5555165°C, 10 min16
FastDigest BplI37°C15
(lambda DNA/XhoI)
203030nd65°C, 5 min16
FastDigest Bpu10I37°C15
(M13mp18 DNA)
153015380°C, 5 min1
FastDigest Bpu1102I37°C5555280°C, 5 min16
FastDigest BseDI37°C5555380°C, 5 min16
FastDigest BseGI37°C5555280°C, 5 min16
FastDigest BseJI65°C55552No (chloroform extraction)1
FastDigest BseLI37°C55552No (chloroform extraction)16
FastDigest BseMI37°C15151515380°C, 5 min16
FastDigest BseMII55°C15301515280°C, 5 min16
FastDigest BseNI65°C5555280°C, 5 min16
FastDigest BseSI37°C55530380°C, 15 min16
FastDigest BseXI65°C15
(pBR322 DNA)
151515380°C, 15 min16
FastDigest Bsh1236I37°C5555180°C, 10 min16
FastDigest Bsh1285I37°C15151515380°C, 15 min1
FastDigest BshNI37°C560515265°C, 10 min16
FastDigest BshTI37°C5555380°C, 5 min16
FastDigest Bsp119I37°C5555180°C, 5 min16
FastDigest Bsp120I37°C55155380°C, 10 min16
FastDigest Bsp1407I37°C5555380°C, 10 min16
FastDigest Bsp143I37°C55105465°C, 20 min16
FastDigest BspLI37°C5555265°C, 20 min16
FastDigest BspOI37°C55155380°C, 5 min1
FastDigest BspTI37°C55554No (chloroform extraction)16
FastDigest Bst1107I37°C555103No (chloroform extraction)6
FastDigest BstXI37°C5555480°C, 5 min1
FastDigest Bsu15I37°C5555265°C, 15 min16
FastDigest BsuRI37°C55553No (chloroform extraction)16
FastDigest BveI37°C15156020565°C, 5 min16
FastDigest CaiI37°C5555465°C, 10 min16
FastDigest Cfr10I37°C551052No (chloroform extraction)2
FastDigest Cfr13I37°C55552No (chloroform extraction)16
FastDigest CpoI37°C5555165°C, 5 min16
FastDigest CseI37°C15
(pBR322 DNA)
153015280°C, 5 min16
FastDigest CsiI37°C510515465°C, 5 min16
FastDigest Csp6I37°C5555280°C, 10 min16
FastDigest DpnI37°C5
(pBR322 DNA)
5not applicable10180°C, 5 min16
FastDigest DraI37°C5555465°C, 5 min16
FastDigest Eam1104I37°C5
(phiX174 DNA)
555380°C, 5 min6
FastDigest Eam1105I37°C5555365°C, 5 min16
FastDigest Ecl136II37°C5555165°C, 5 min6
FastDigest Eco105I37°C5555365°C, 5 min16
FastDigest Eco130I37°C552020365°C, 5 min16
FastDigest Eco147I37°C5555380°C, 10 min16
FastDigest Eco31I37°C5555365°C, 5 min16
FastDigest Eco32I37°C55552No (chloroform extraction)16
FastDigest Eco47I37°C5555180°C, 20 min16
FastDigest Eco47III37°C5555465°C, 5 min16
FastDigest Eco52I37°C520205365°C, 5 min16
FastDigest Eco57I37°C15301515265°C, 5 min16
FastDigest Eco72I37°C5555280°C, 10 min1
FastDigest Eco81I37°C5555280°C, 10 min16
FastDigest Eco88I37°C5555265°C, 5 min16
FastDigest Eco91I37°C5555265°C, 10 min2
FastDigest EcoO109I37°C510155265°C, 5 min16
FastDigest EcoRI37°C55205280°C, 5min0.5
FastDigest EheI37°C5555265°C, 5 min6
FastDigest Esp3I37°C5555165°C, 10 min6
FastDigest FaqI37°C15301515365°C, 5 min16
FastDigest FokI37°C5555165°C, 5 min1
FastDigest FspAI37°C5555465°C, 5 min16
FastDigest FspBI37°C515520380°C, 5 min16
FastDigest GsuI30°C15301515265°C, 5 min16
FastDigest HhaI37°C55551No (chloroform extraction)16
FastDigest Hin1I37°C5555165°C, 5 min16
FastDigest Hin1II37°C510510480°C, 5 min16
FastDigest Hin6I37°C51055280°C, 10 min16
FastDigest HincII37°C55510165°C, 5 min16
FastDigest HindIII37°C552010380°C, 10 min16
FastDigest HinfI37°C5555265°C, 20 min16
FastDigest HpaII37°C5555365°C, 5 min16
FastDigest Hpy8I37°C55510280°C, 5 min16
FastDigest HpyF10VI37°C5555280°C, 5 min16
FastDigest HpyF3I37°C55510365°C, 5 min16
FastDigest KflI37°C5
55105No (chloroform extraction)16
FastDigest Kpn2I37°C5555280°C, 5 min16
FastDigest KpnI37°C5555380°C, 5 min16
FastDigest KspAI37°C5555365°C, 20 min0.5
FastDigest LguI37°C5555265°C, 5 min16
FastDigest Lsp1109I37°C5
(pBR322 DNA)
5510265°C, 5 min1
FastDigest MauBI37°Cno sites
(pJET1-MauBI DNA/BamHI, 5 min)
5105565°C, 5 min16
FastDigest MbiI37°C5555365°C, 5 min16
FastDigest MboI37°C55510165°C, 15 min16
FastDigest MboII37°C5555265°C, 5 min16
FastDigest MlsI37°C553020380°C, 20 min16
FastDigest MluI37°C55515380°C, 5 min16
FastDigest MnlI37°C5555265°C, 5 min16
FastDigest Mph1103I37°C55510365°C, 15 min6
FastDigest MreI37°Cno sites
(pJET1-MreI DNA/Eam1105I, 5 min)
555380°C, 5 min16
FastDigest MspI37°C55553No (chloroform extraction)16
FastDigest MssI37°C5555265°C, 10 min16
FastDigest MunI37°C55553No (chloroform extraction)16
FastDigest Mva1269I37°C5555365°C, 5 min16
FastDigest MvaI37°C555104No (chloroform extraction)1
FastDigest NcoI37°C510105365°C, 15 min16
FastDigest NdeI37°C556030365°C, 5 min6
FastDigest NheI37°C51555565°C, 5 min6
FastDigest NmuCI37°C55510265°C, 5 min16
FastDigest NotI37°Cno sites
(pTZ19RJL2 DNA/BseLI, 5 min)
30510280°C, 5 min16
FastDigest NsbI37°C5555365°C, 15 min16
FastDigest OliI37°C5
(phiX174 DNA)
5105365°C, 5 min6
FastDigest PacI37°Cno sites
(pJET1-PacI DNA/SdaI, 5 min)
555265°C, 10 min16
FastDigest PaeI37°C5555565°C, 5 min16
FastDigest PagI37°C5
(phiX174 DNA)
10510380°C, 5 min0.5
FastDigest PdiI37°C5
(pBR322 DNA/NdeI)
555465°C, 15 min16
FastDigest PdmI37°C5555265°C, 5 min16
FastDigest PfeI37°C55105265°C, 5 min16
FastDigest Pfl23II37°C15151515365°C, 5 min16
FastDigest PfoI37°C5555365°C, 5 min16
FastDigest Ppu21I37°C5555165°C, 5 min16
FastDigest PsyI37°C5555280°C, 5 min4
FastDigest Psp1406I37°C5555365°C, 5 min16
FastDigest Psp5II37°C55520280°C, 5 min6
FastDigest PspFI37°C5555480°C, 5 min16
FastDigest PstI37°C553053No (chloroform extraction)16
FastDigest PsuI37°C5555280°C, 5 min16
FastDigest PteI37°C5555380°C, 5 min16
FastDigest PvuI37°C51555480°C, 5 min16
FastDigest PvuII37°C55551No (chloroform extraction)0.5
FastDigest RruI37°C55555No (chloroform extraction)4
FastDigest RsaI37°C55553No (chloroform extraction)16
FastDigest RseI37°C5555465°C, 20 min16
FastDigest SacI37°C5153010165°C, 5 min16
FastDigest SalI37°C55605365°C, 10 min16
FastDigest SaqAI37°C5
(pBR322 DNA)
555465°C, 5 min1
FastDigest SatI37°C55105365°C, 5 min16
FastDigest ScaI37°C5555465°C, 10 min16
FastDigest SchI37°C55305280°C, 5 min1
FastDigest SdaI37°C55553No (chloroform extraction)1
FastDigest SduI37°C5555280°C, 15 min1
FastDigest SfaAI37°Cno sites
(pJET1-AsiSI DNA/SdaI, 5 min)
5605580°C, 5 min16
FastDigest SfiI50°Cno sites
(pUC19-SfiI DNA, 15 min)
1515155No (chloroform extraction)16
FastDigest SgsI37°C5555265°C, 20 min16
FastDigest SmaI37°C5555165°C, 5 min16
FastDigest SmiI37°Cno sites
(M13mp18 DNA/MvaI, 5 min)
203030265°C, 15 min1
FastDigest SsiI37°C5555265°C, 5 min4
FastDigest SspI37°C55530265°C, 5 min1
FastDigest TaaI65°C55553No (chloroform extraction)16
FastDigest TaiI65°C55552No (chloroform extraction)16
FastDigest TaqI65°C55554No (chloroform extraction)16
FastDigest TasI65°C5
(pBR322 DNA)
5553No (chloroform extraction)6
FastDigest TatI65°C151515205No (chloroform extraction)1
FastDigest TauI55°C156015303No (chloroform extraction)16
FastDigest Tru1I65°C555103No (chloroform extraction)2
FastDigest TscAI65°C55552No (chloroform extraction)6
FastDigest Van91I37°C5555365°C, 10 min6
FastDigest VspI37°C55530265°C, 5 min16
FastDigest XagI37°C55520265°C, 5 min16
FastDigest XapI37°C55510480°C, 5 min16
FastDigest XbaI37°C55510265°C, 20 min16
FastDigest XceI37°C5555265°C, 5 min16
FastDigest XhoI37°C55510280°C, 5 min16
FastDigest XmaJI37°C5105102No (chloroform extraction)16
FastDigest XmiI37°C55605265°C, 5 min16


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