Recently Viewed

Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

EcoRII

5'...C C W G G ...3'
3'... G G W C C...5'

O buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by Dcm methylation Thermal inactivation at 80°C in 20 min Recombinant enzyme LO certified

The EcoRII restriction enzyme recognizes ^CCWGG sites and cuts best at 37°C in O buffer (Isoschizomers: AjnI, BseBI, Bst2UI, BstNI, BstOI, Psp6I, PspGI)
  
Loading...
Lambda DNA (dcm-) digested with EcoRII

Lambda DNA (dcm-) digested with EcoRII, 1.4% agarose, 71 cleavage sites

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

  • At least two copies of EcoRII recognition site are required for efficient cleavage. For cleavage of DNA substrates with only one copy of recognition site MvaI, neoschizomer of EcoRII is recommended.
  • EcoRII may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation, or heat the digested DNA in the presence of SDS prior to electrophoresis.
  • Assayed using pBR322 DNA (dcm-). EcoRII is blocked by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsBssKI - (CCWGG), PfoI - (TCCWGGA), SexAI.
Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferEcoRII is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X
G (green)
1X
O (orange)
1X
R (red)
1X
Tango
(yellow)
1X / 2X
O (Orange) 37°C 20-50 50-100 100 50-100 20-50 50-100 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: completely overlaps - blocked.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dcm (CCWGG)Cm5CWGG Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0 0-20 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
71 2 7 6 5 5
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
5 5 5 5 8 12

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
^CCAGG
  • FastDigest SexAI (CsiI)* (A^CCAGGT)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BssKI
  • EcoRII
  • MvaI (BstNI)/FastDigest MvaI
  • SetI
^CCTGG
  • FastDigest SexAI (CsiI)* (A^CCTGGT)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BssKI
  • EcoRII
  • MvaI (BstNI)/FastDigest MvaI

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
^CCWGGCCWGGCCWGG
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • [EcoRII]
  • [MvaI (BstNI)/FastDigest MvaI]
^CCAGGCCAGGCCAGG
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • [EcoRII]
  • [MvaI (BstNI)/FastDigest MvaI]
^CCTGGCCTGGCCTGG
  • [Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)]
  • [BssKI]
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • [EcoRII]
  • [MvaI (BstNI)/FastDigest MvaI]