FastDigest Enzymes

176 FastDigest enzymes.

1 buffer.

Overview

View or download the FastDigest product brochure

Thermo Scientific FastDigest enzymes are an advanced line of restriction enzymes that create a new standard in 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 large section of enzyme isoschizomers and state-of-the-art production facility facilitated the creation of the unique system of 176 FastDigest® restriction enzymes.

Old Standard
Conventional restriction enzymes
New Standard
FastDigest restriction enzymes
Buffer System Up to 20 Buffers One Universal Buffer
Double/Multiple Restriction Digestion Limited by buffer compatibility Any enzyme combination possible
Reaction Time 1 hour - overnight 5 - 15 minutes
Direct Loading On Gels No Yes
Downstream Applications Partial compatibility 100% buffer compatibility
Star Activity Yes No
Activity Definintion 1 unit of enzyme hydrolyzes 1µg of lambda DNA in 60 minutes in an optimal buffer for an enzyme 1 µL of FastDigest enzyme cleaves1 µg of substrate DNA in 5 to 15 minutes in FastDigest buffer

100% activity in a single buffer

The FastDigest and FastDigest Green Buffer is a proprietary digestion buffer which supports 100% activity of all FastDigest restriction enzymes.

Benefits:

  • Double and multiple digestion in a single buffer with any combination of enzymes.
  • No sequential digestions or buffer changes needed.
  • Allows direct loading of reaction mixture on gels.
  • Saves pipetting time and eliminates the need for DNA loading dyes.
FastDigest Buffer and FastDigest Green Buffer

Five minute triple digestion of plasmid DNA followed by ligation in FastDigest and FastDigest Green buffers

  • M:GeneRuler™ Express DNA Ladder (#SM1551)
  • C:Undigested plasmid DNA control
  • 1:Plasmid DNA triple digested with FastDigest EcoRI, FastDigest KpnI and FastDigest SmaI in FastDigest buffer
  • 2:The digestion and ligation reaction mixture in FastDigest buffer
  • 3:Plasmid triple digested with FastDigest EcoRI, FastDigest KpnI and FastDigest SmaI in FastDigest Green buffer
  • 4:The digestion and ligation reaction mixture in FastDigest Green buffer

100% buffer compatibility with downstream applications

DNA/RNA modifying enzymes, such as ligases, phosphatases, kinases and mesophilic DNA polymerases have 100% activity in FastDigest and FastDigest Green Buffer. Therefore, enzymes used in downstream applications can be directly added to the FastDigest reaction mix.

DNA/RNA Modifying Enzyme Activity in FastDigest Green Buffer/FastDigest Buffer
DNA Polymerase I, E.coli 100%
Klenow Fragment 100%
Klenow Fragment, exo 100%
T4 DNA Polymerase 100%
T7 DNA Polymerase 100%
T4 DNA Ligase* 75-100%
FastAP Thermosensitive Alkaline Phosphatase 100%
T4 Polynucleotide Kinase 100%
* 0.5 mM ATP is required for T4 DNA Ligase activity.

Direct loading on gels

As an added convenience we developed the FastDigest Green Buffer which offers the same performance as the colorless FastDigest Buffer but enables direct loading of the reaction mixture on gels. The 10X FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading. 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.

FastDigest Green Buffer

FastDigest Green Buffer

Reaction mixture containing FastDigest Green Buffer:

  1. Loaded into gel well, before electrophoresis
  2. Separated, after electrophoresis

No star activity


FastDigest enzymes are designed to eliminate star activity:
  • Short incubation time (5-15 min), no need for prolonged digestions.
  • Universal buffer with optimal pH and salt concentration for all enzymes.
  • Optimal enzyme and glycerol concentrations.

Conventional restriction enzymes may display star or "relaxed" activity due to prolonged incubation times, high enzyme and/or glycerol concentration, high pH values or low ionic strength. By addressing all these issues, FastDigest enzymes enable single, double and even triple digestion in 5 minutes without any signs of star activity.

FastDigest No Star Activity

Five minute single and double digestion of plasmid DNA with no signs of star activity

  • M:GeneRuler 1 kb Plus DNA Ladder (#SM1333)
  • C: Undigested plasmid DNA
  • 1:Plasmid DNA digested in 5 min with FastDigest SmaI
  • 2:Plasmid DNA digested in 5 min with FastDigest EcoRI
  • 3:Plasmid DNA double digested in 5 min with FastDigest SmaI and FastDigest EcoRI

Complete digestion in 5-15 minutes


Double and multiple digestions in one buffer in 5-15 minutes:
  • Saves time and effort, increasing throughput.
  • Digestion times are provided for all types of DNA templates (plasmid DNA, PCR product, genomic DNA).
  • No overnight digestions are required for any template.
  • Star activity is eliminated due to short reaction times.
Conventional restriction enzymes FastDigest restriction enzymes
Sequential digestion One reaction mixture
Reaction setup for Apal ~2 min
Reaction set up with FastDigest ApaI and XhoI ~2 min
Incubation 60 min
Incubation 5 min
Reaction setup for Xhol ~2 min
Incubation 60 min
TOTAL TIME >2 hours
TOTAL TIME 7 min

176 FastDigest enzymes available

During 30 years of restriction enzyme search program we acquired one of the largest collections of restriction enzyme producing bacterial strains in the industry. Our wide selection of isoschizomers and ability to produce enzymes of exceptional purity enabled us to create the unique system of 176 FastDigest restriction enzymes formulated to have 100% activity in the universal buffer.

30 years of experience
>3000 restriction enzyme producing strains
>30% of all known enzymes discovered
>30% of all known enzymes cloned
176 Thermo Scientific FastDigest Enzymes available
FastDigest Green Buffer for direct loading on gels

RReaction Conditions for FastDigest Enzymes Chart  
Digestion of Methylated DNA
Buffer Compatibility for Modifying Enzymes  
Available On-Site