FastDigest—Thermo Scientific
One buffer for 176 enzymes. It's that easy.
Thermo Scientific FastDigest Restriction Enzymes are an advanced line of enzymes offering fast and complete digestion of DNA in a single universal buffer. With over 30 years experience in support of restriction enzyme research, our FastDigest enzymes are a top customer pick offering 176 unique specificities in formulations that support direct loading of reaction mixtures on gels.
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176 FastDigest enzymes available
Over the course of 30 years, our search for restriction enzymes led us to acquire 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 a single buffer.
30 years of experience
- >3000 restriction enzyme producing strains
- >30% of all known enzymes discovered
- >30% of all known enzymes cloned
- 176 FastDigest enzymes available
- Thermo Scientific FastDigest Green Buffer for direct loading on gels
| Conventional restriction enzymes | 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 Green Buffer and Thermo Scientific FastDigest Buffer are proprietary digestion buffers which support 100% activity of all FastDigest restriction enzymes. This system allows for double and multiple digestions with any combination of enzymes. No sequential digestions or buffer changes are needed.
Five minute single, double and triple digestions in FastDigest Green Buffer
Five minute plasmid DNA digestions with:
- Lane M: Thermo Scientific GeneRuler Express Ladder (Cat. No. SM1553)
- Lane 1: Control, undigested plasmid DNA
- Lane 2: FastDigest EcoRI
- Lane 3: FastDigest EcoRI and FastDigest KpnI
- Lane 4: FastDigest EcoRI, FastDigest KpnI and FastDigest SmaI
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. No buffer changes or purification steps are needed.
| 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.
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 Sequential digestion |
FastDigest restriction enzymes 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 |
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
Reaction mixture containing FastDigest Green Buffer:
- Lane 1: Loaded into gel well, before electrophoresis
- Lane 2: Separated, after electrophoresis
Short protocol times with no star activity
FastDigest enzymes in combination with FastDigest (Green) Buffer 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.
Five minute single and double digestion of plasmid DNA with no signs of star activity
Five minute plasmid DNA digestions with:
- Lane 1: FastDigest SmaI
- Lane 2: FastDigest EcoRI
- Lane 3: FastDigest SmaI and FastDigest EcoRI double digest
- Lane M: GeneRuler 1 kb Plus DNA Ladder (Cat. No. SM1333)
- Lane C: Control, Undigested plasmid DNA
Resources
For Research Use Only. Not for use in diagnostic procedures.
