Introduction: Restriction Enzymes
Description

Restriction enzymes (or restriction endonuclease) are enzymes that cut double-stranded DNA at their specific recognition nucleotide sequences known as restriction sites. In nature, they function as microbial immune systems against invading viruses. Restriction enzyme digestion has been a routine method of molecular biology for more than 2 decades. Until now researches use restriction enzymes for cloning, analysis of genomic sequences (RFLP) and DNA methylation. The application of restriction enzymes has made everything in genetic engineering and recombinant DNA technology possible.

In year 2011, Gentaur begins to offer 78 high-quality restriction enzymes for our customer all over the world.
Features

  • 100% buffer compatibility with downstream applications
  • Complete digestion in 10-30 minutes
  • 73 enzymes available
  • 25 high concentration formats available
  • Different units, same price
Applications

  • Cloning and routine gene manipulation
  • Analysis of genomic sequences (RFLP)
  • DNA methylation
Unit Definition

One unit is defined as the amount of enzyme required to digest 1 μg of λ DNA in 60 min at 37°C in a reaction volume of 50 μl.
Quality Control

5-Hour Incubation:
Incubation of 20 units of each restriction enzyme with 1 μg of DNA subtract for 5 hours (100-fold over-digestion) at 37°C resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

Ligation and Re-cutting:
λDNA was 4-fold over-digested with each restriction enzyme for 2 hours. The DNA fragments were then ligated with T4 DNA ligase. The ligated DNA could be recut with the enzyme. The DNA ligation and re-cutting patterns were determined by agarose gel electrophoresis.
Quality Control

Endonuclease activity:
Incubation of 10-35 units of each restriction enzyme with 1 μg of ΦX174 or pBR322 RF I DNA for 5 hours at 37°C resulted in < 10% conversion to RFII as determined by agarose gel electrophoresis. 

Phosphatase activity:
Incubation of a 20-200 units of each restriction enzyme with p-nitrophenyl phosphate for 48 hours at 37°C released < 0.1% of free p-nitrophenol.
Restriction Enzymes Buffer Map

Buffer Information (1X):

 

Inactivation Temperature Table

 

Storage and Shipping conditions

All restriction enzymes should be stored at -20°C. Enzymes typically are shipped on dry ice. During shipment they may freeze. However, this does not affect their quality. Enzymes may tolerate few times of freeze-thaw cycles without losing any activity. However, frequent freeze-thaw cycles will decrease enzymatic activity gradually. For enzymes in large packages, we recommend our customers to aliquot enzymes immediately once arrival.
Specification
Store at

 

All restriction enzymes should be stored at -20°C.

 

Additonal Starter Pack
# FYR000-5RE

BamH I 1000 U
Hind III 1000 U
EcoR I 1000 U
Xba I 1000 U
Sma I 500 U
# FYR000-1M-13RE
BamH I 1000 U
Hind III 1000 U
EcoR I 1000 U
Xba I 1000 U
Sma I 500 U
Pst I 800 U
EcoR V 200 U
Kpn I 200 U
Sac I 200 U
Sal I 220 U
Xho I 500 U
Not I 50 U
Bgl II 200 U
# FYD002-500UL
(100 bp YEA Ladder DNA Marker)
30 μg/500 μl
# FYR000-2M-13RE
BamH I 1000 U
Hind III 1000 U
EcoR I 1000 U
Xba I 1000 U
Sma I 500 U
Pst I 800 U
EcoR V 200 U
Kpn I 200 U
Sac I 200 U
Sal I 220 U
Xho I 500 U
Not I 50 U
Bgl II 200 U
100 bp YEA Ladder DNA Marker 15 μg/250 μl
1Kb YEA Ladder DNA Marker 10 μg/250 μl
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