176 FastDigest enzymes.

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.

The 10X FastDigest Green Buffer is a proprietary digestion buffer which supports 100% activity of all FastDigest restriction enzymes.

Benefits:

• Double and multiple digestion in the universal 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.

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

DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase have 100% activity in FastDigest Green Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

The 10X FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products 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.

FastDigest Green Buffer

Reaction mixture containing FastDigest Green Buffer:

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

• 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

• M:GeneRuler 1 kb Plus DNA Ladder
• 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

• 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.

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.