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

CseI (HgaI)

5'...G A C G C (N)5 ↓...3'
3'...C T G C G (N)10↑...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available R buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Only small amounts (up to 10 u) can be inactivated at 80°C in 20 min Recombinant enzyme LO certified

The CseI (HgaI) restriction enzyme recognizes GACGC(5/10)^ sites and cuts best at 37°C in R buffer (Isoschizomers: HgaI)
  
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pBR322 DNA digested with CseI (HgaI)

pBR322 DNA digested with CseI (HgaI), 1.4% agarose, 11 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

Assayed using pBR322 DNA (#SD0041). Low salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity. CseI 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.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer R: 10 mM Tris-HCl (pH 8.5 at 37°C), 10 mM MgCl2, 100 mM KCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.

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
R (Red) 37°C NR 50-100† 50-100 100 100† 50-100 1X† or 2X

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - no effect.
  • CpG: completely overlaps - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
CpGGACGC Blocked

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
102 14 7 11 4 4
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
4 4 4 4 5 9