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.


FastDigest restriction digest finder by enzyme, buffer, or optimal temperature

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
(pJET1-SanDI DNA/BamHI)
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

Note

  • (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.

For Research Use Only. Not for use in diagnostic procedures.

Stylesheet for Classic Wide Template adjustments