Reaction Conditions for FastDigest Enzymes | Thermo Fisher Scientific

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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
Thermo Scientific FastDigest enzyme	Reaction temperature	Lambda, 1 µg/20 µL	Plasmid DNA, 1 µg/20 µL	PCR product, ~0.2 µg/30 µL	Genomic DNA, 1 µg/10 µL	bp from end of DNA required for complete digestion	Thermal inactivation	Incubation time (in hours) without star activity
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

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.