1 buffer. 176 enzymes. 100% activity. - thermofisher.com · advanced line of restriction enzymes...
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1 buffer. 176 enzymes. 100% activity.Restriction digestion as easy as it gets
• Complete digestion in 5-15 minutes• 100% buffer compatibility with downstream applications• Direct loading on gels
Thermo Scientific FastDigest Restriction Enzymes
Features
100% activity of all FastDigest enzymes in one buffer
Complete digestion in 5-15 minutes
100% buffer compatibility with downstream applications
Direct loading on gels
176 FastDigest enzymes available
ApplicationsClone analysis
Preparation of DNA for cloning
Digestion of PCR products
RFLP genotyping
Digestion of difficult-to-cleave DNA
All FastDigest enzymes are 100% active in the universal Thermo Scientific™ FastDigest™ and FastDigest Green buffers and are able to digest DNA in 5-15 minutes. This enables any combination of restriction enzymes to work simultaneously in one reaction tube and eliminates the need for sequential digestions. In addition the FastDigest Green Buffer allows for direct loading of the reaction mixture on gels. FastDigest enzymes are qualified for complete digestion of lambda DNA, plasmid DNA, genomic DNA, and PCR products. Digestion time and protocols are experimentally tested and provided for each FastDigest enzyme on each of the templates.
Thermo Scientific™ FastDigest™ enzymes are an
advanced line of restriction enzymes for rapid DNA
digestion. During our 30 years of restriction enzyme
research, we compiled one of the largest collections
of restriction enzyme producing bacterial strains in the
industry. Our profound expertise, the large selection of
enzyme isoschizomers and state-of-the-art production
facilities led to the creation of the first simplified
restriction enzyme system using a universal buffer.
Restriction digestion as easy as it gets
* 0.5 mM ATP is required for T4 DNA Ligase activity.
DNA/RNA modifying enzymeActivity in FastDigest Green Buffer/
FastDigest Buffer
DNA Polymerase I, E.coli 100%
Klenow Fragment 100%
Klenow Fragment, exo¯ 100%
T7 DNA Polymerase 100%
T4 DNA Ligase* 75-100%
Thermo Scientifc™ FastAP™ Thermosensitive Alkaline Phosphatase
100%
T4 Polynucleotide Kinase 100%
100% buffer compatibility with all downstream applications
Common DNA modifying enzymes are 100% active in FastDigest and FastDigest Green Buffer.
Therefore, enzymes used in downstream applications can be added directly to the restriction digestion reaction mixture, eliminating the need for additional DNA purification steps.
More than 35 years of experience in the development of restriction enzymes
Fermentas founded
176 FastDigest enzymes available
>3000 restriction enzyme producing
strains
18 unique restriction enzymes available
>40% of all known restriction enzymes
cloned
First unique enzyme
discovered, CfrIWorld’s first “artificial” enzymes Eco57MI and
Nb.Bpu10I created
First restriction enzyme cloned,
Eco47IILaunch of FastDigest
line
>30% of all known restriction enzymes characterized
Acquisition by Thermo Fisher
Scientific
OUR EXPERTISE TODAY
Launch of FastDigest
Green Buffer
1975 1977 1985 2001 2006 2009 2010
Set a new standard in DNA digestion
OLD STANDARDConventional restriction enzymes
NEW STANDARDFastDigest restriction enzymes
Buffer system Up to 20 buffers One universal buffer
Buffer change between enzymatic steps Frequently necessary Not necessary
Double/multiple restriction digestion Limited due to buffer incompatibility Unlimited – all 176 enzymes are 100% active in one buffer
Reaction time for plasmid DNA, genomic DNA, PCR product
1 hour-overnight 5-15 minutes
Direct loading of reaction mixture on gels No Yes
Compatibility with downstream applications
Partial 100%
Star activity Dependent on enzyme properties and reaction conditions
No star activity due to short incubation time
Activity definition 1 unit of enzyme hydrolyzes 1 μg of lambda DNA in 60 minutes in an optimal buffer for an enzyme
1 μL of FastDigest enzyme cleaves 1 μg of substrate DNA in 5 or 15 minutes in FastDigest buffer
Universal buffer and direct loading on gels
Our FastDigest line of restriction enzymes are ideal for use in applications that require high-purity reaction components, performance reliability and simple reaction set-up.
As an added convenience we developed the FastDigest Green Buffer, which offers the same performance as the colorless FastDigest buffer. All FastDigest restriction enzymes are 100% active in FastDigest Green Buffer. The green buffer contains a density reagent and two tracking dyes to enable direct loading of reaction mixture on gels. The blue dye migrates with 3-5 kb DNA fragments in a 1% agarose gel and has an excitation peak of 424 nm. The yellow dye migrates faster than 10 bp DNA fragments in a 1% agarose gel and has an excitation peak of 615 nm.
The FastDigest Green Buffer exhibits the same high efficiency in DNA digestion and downstream applications as the colorless FastDigest Buffer. Only for applications that require product analysis by fluorescence excitation (e.g., concentration measurements in UV light), we recommend using the colorless 10X FastDigest Buffer.
Highlights
Double and multiple digestion in one buffer for any combination of enzymes
No sequential digestions and buffer changes
100% buffer compatibility with downstream applications
Direct loading of reaction mixtures on gels saves sample loading reagents and time
Highest quality and performance
Just like all Thermo Scientific™ Molecular Biology enzymes and kits, the quality and performance of FastDigest enzymes benefit from an industry leading manufacturing process. Production is taking place in Class D clean room facilities that
are qualified and certified as per EU directives and ISPE guidelines. We apply stringent quality control testing and assure consistency and lot-to-lot reproducibility in accordance with ISO9001 and ISO14001.
Conventional restriction enzymesSequential digestion
FastDigest restriction enzymesOne reaction mixture
Reaction setup for ApaI ~ 2 minutes
Incubation at 37°C 60 minutes
Reaction setup for Xhol > 2 minutes
Incubation at 37°C 60 minutes
1 Setup ONE reaction mixture with: FastDigest ApaI and FastDigest XhoI ~2 minutes
2 Incubation at 37°C 5 minutes
1 1
22
3
4
TOTAL TIME > 2 hours
TOTAL TIME 7 minutes
Save more than 2 hours when performing double digestion reactions
5 minute plasmid DNA digestions with: 1: FastDigest EcoRI2: FastDigest EcoRI and FastDigest KpnI3: FastDigest EcoRI, FastDigest KpnI and FastDigest SmaIM: Thermo Scientific™ GeneRuler™ Express Ladder (#SM1553) C: Undigested plasmid DNA
Five minute single, double and triple digestions in FastDigest Green Buffer
M C 1 2 3 M
Double and multiple digestion in one buffer in 5-15 minutes
Digestion times provided for all types of DNA templates: plasmid DNA, PCR product, genomic DNA
No overnight digestions for any template
No star activity due to shortened incubation times
Ideal for high-throughput applications
Rapid DNA digestion with FastDigest enzymes shows the same results as longer digestion with conventional restriction enzymes. 5 min plasmid DNA digestion with:1: FastDigest XhoI, 2: FastDigest ApaI3: FastDigest XhoI and FastDigest ApaI1 hour plasmid DNA digestion with4: ApaI 5: XhoI6: Sequential double digestion with ApaI (1x Tango buffer, 4-fold excess of
ApaI, 1 hour) and XhoI (2x Tango buffer, 1 hour)M: Thermo Scientific™ GeneRuler™ 1 kb Plus DNA Ladder (#SM1334)C: Undigested plasmid DNA
M C 1 2 3 M C 4 5 6 M
5 Min5 Min
5 Min
60 min60 min 120 min
FastDigest enzymes Conventional enzymes
Comparison of FastDigest and conventional restriction enzymes
DNA digestion in 5-15 minutes
Reaction mixture containing FastDigest Green Buffer:1: Loaded into gel well,
before electrophoresis2: Separated, after
electrophoresis10 bp
3-5 kb
1 2
FastDigest Green Buffer
Order information
Cat. No. Description Rxns.FD2064 FastDigest AanI 20FD1724 FastDigest AasI 25FD0994 FastDigest AatII 50FD0904 FastDigest Acc65I 100FD1234 FastDigest AdeI 50FD1954 FastDigest AjuI 20FD0014 FastDigest AluI 100FD0024 FastDigest Alw21I 100FD0034 FastDigest Alw26I 100FD0044 FastDigest Alw44I 200FD1414 FastDigest ApaI 300FD0054/5 FastDigest BamHI 800/2500FD0724 FastDigest BclI 300FD0064 FastDigest BcnI 200FD1253/4 FastDigest BcuI 20/50FD1164 FastDigest BfmI 20FD2184 FastDigest BfoI 200FD0074 FastDigest BglI 200FD0083/4 FastDigest BglII 100/200FD1424 FastDigest Bme1390I 100FD2124 FastDigest BmsI 20FD1434 FastDigest BoxI 100FD1014 FastDigest BpiI 20FD1314 FastDigest BplI 20FD1184 FastDigest Bpu10I 20FD0094 FastDigest Bpu1102I 50FD1084 FastDigest BseDI 20FD0874 FastDigest BseGI 100FD1714 FastDigest BseJI 200FD1204 FastDigest BseLI 100FD1264 FastDigest BseMI 20FD1404 FastDigest BseMII 20FD0884 FastDigest BseNI 100FD1444 FastDigest BseSI 50FD1454 FastDigest BseXI 20FD0924 FastDigest Bsh1236I 100FD0894 FastDigest Bsh1285I 100FD1004 FastDigest BshNI 300FD1464 FastDigest BshTI 20FD0124 FastDigest Bsp119I 200FD0134 FastDigest Bsp120I 200FD0933/4 FastDigest Bsp1407I 50/100FD0784 FastDigest Bsp143I 40FD1154 FastDigest BspLI 20
Cat. No. Description Rxns.FD2044 FastDigest BspOI 20FD0834 FastDigest BspTI 150FD0704 FastDigest Bst1107I 100FD1024 FastDigest BstXI 100FD0143/4 FastDigest Bsu15I 50/100FD0154 FastDigest BsuRI 400FD1744 FastDigest BveI 50FD1394 FastDigest CaiI 50FD0184 FastDigest Cfr10I 50FD0194 FastDigest Cfr13I 100FD0744 FastDigest CpoI 20FD1904 FastDigest CseI 20FD2114 FastDigest CsiI 50FD0214 FastDigest Csp6I 100FD1703/4 FastDigest DpnI 50/100FD0224 FastDigest DraI 200FD0234 FastDigest Eam1104I 50FD0244 FastDigest Eam1105I 100FD0254 FastDigest Ecl136II 100FD0404 FastDigest Eco105I 50FD0414 FastDigest Eco130I 200FD0424 FastDigest Eco147I 100FD0293/4 FastDigest Eco31I 50/100FD0303/4 FastDigest Eco32I 200/400FD0314 FastDigest Eco47I 200FD0324 FastDigest Eco47III 20FD0334 FastDigest Eco52I 50FD0344 FastDigest Eco57I 20FD0364 FastDigest Eco72I 200FD0374 FastDigest Eco81I 50FD0384 FastDigest Eco88I 200FD0394 FastDigest Eco91I 200FD0264 FastDigest EcoO109I 200FD0274/5 FastDigest EcoRI 800/2500FD0443/4 FastDigest EheI 20/50FD0454 FastDigest Esp3I 20FD1814 FastDigest FaqI 20FD2144 FastDigest FokI 100FD1664 FastDigest FspAI 20FD1764 FastDigest FspBI 50FD0464 FastDigest GsuI 20FD1854 FastDigest HhaI 200FD0474 FastDigest Hin1I 100FD1834 FastDigest Hin1II 100
Cat. No. Description Rxns.FD0484 FastDigest Hin6I 200FD0494 FastDigest HincII 100FD0504/5 FastDigest HindIII 800/2500FD0804 FastDigest HinfI 400FD0514 FastDigest HpaII 200FD1574 FastDigest Hpy8I 20FD1734 FastDigest HpyF10VI 20FD1884 FastDigest HpyF3I 50FD2164 FastDigest KflI 20FD0534 FastDigest Kpn2I 100FD0524 FastDigest KpnI 300FD1034 FastDigest KspAI 50FD1934 FastDigest LguI 20FD2074 FastDigest Lsp1109I 100FD2084 FastDigest MauBI 20FD1274 FastDigest MbiI 100FD0814 FastDigest MboI 50FD0824 FastDigest MboII 50FD1214 FastDigest MlsI 50FD0564 FastDigest MluI 100FD1074 FastDigest MnlI 50FD0734 FastDigest Mph1103I 100FD2024 FastDigest MreI 20FD0544 FastDigest MspI 400FD1344 FastDigest MssI 100FD0753/4 FastDigest MunI 20/50FD0964 FastDigest Mva1269I 50FD0554 FastDigest MvaI 200FD0573/4/5 FastDigest NcoI 20/100/300FD0583/4/5 FastDigest NdeI 100/300/1000
FD0973/4 FastDigest NheI 50/100FD1514 FastDigest NmuCI 20FD0593/4/6 FastDigest NotI 20/50/250FD1224 FastDigest NsbI 50FD1634 FastDigest OliI 20FD2204 FastDigest PacI 25FD0604 FastDigest PaeI 20FD1284 FastDigest PagI 50FD1524 FastDigest PdiI 50FD1534 FastDigest PdmI 100FD1784 FastDigest PfeI 50FD0854 FastDigest Pfl23II 50FD1754 FastDigest PfoI 20FD1974 FastDigest Ppu21I 50
Cat. No. Description Rxns.FD0944 FastDigest Psp1406I 20FD0764 FastDigest Psp5II 50FD2224 FastDigest PspFI 20FD0614/5 FastDigest PstI 800/2500FD1554 FastDigest PsuI 150FD1334 FastDigest PsyI 100FD2134 FastDigest PteI 20FD0624 FastDigest PvuI 20FD0634 FastDigest PvuII 400FD2154 FastDigest RruI 100FD1124 FastDigest RsaI 100FD2004 FastDigest RseI 20FD1133/4 FastDigest SacI 100/200FD0644 FastDigest SalI 200FD2174 FastDigest SaqAI 50FD1644 FastDigest SatI 20FD0434 FastDigest ScaI 100FD1374 FastDigest SchI 100FD1194 FastDigest SdaI 20FD0654 FastDigest SduI 50FD2094 FastDigest SfaAI 100FD1824 FastDigest SfiI 150FD1894 FastDigest SgsI 100FD0663/4 FastDigest SmaI 100/200FD1244 FastDigest SmiI 200FD1794 FastDigest SsiI 20FD0774 FastDigest SspI 100FD1364 FastDigest TaaI 20FD1144 FastDigest TaiI 50FD0674 FastDigest TaqI 400FD1354 FastDigest TasI 100FD1294 FastDigest TatI 20FD1654 FastDigest TauI 20FD0984 FastDigest Tru1I 50FD2104 FastDigest TscAI 100FD0714 FastDigest Van91I 100FD0914 FastDigest VspI 200FD1304 FastDigest XagI 100FD1383/4 FastDigest XapI 50/100FD0684/5 FastDigest XbaI 300/750FD1474 FastDigest XceI 50FD0694/5 FastDigest XhoI 400/1200FD1564 FastDigest XmaJI 20FD1484 FastDigest XmiI 50
Set a new standard in DNA digestion
• For further information visit: thermoscientific.com/fastdigest
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Lit # MB1457
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