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SgeI

5'...m5C N N G (N)9 ↓...3'
3'...  G N N C (N)13↓...5'†
† SgeI cleaves DNA targets containing 5-methylcytosine on one or both DNA strands

Unique buffer for 100% activity Optimal incubation at 37°C Star activity Thermal inactivation at 65°C in 20 min Genome qualified Recombinant enzyme LO certified

The SgeI restriction enzyme recognizes m5CNNG(9/13)^ sites and cuts best at 37°C in its own unique buffer
  
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pBR322 DNA (dcm+)digested with SgeI

pBR322 DNA (dcm+) digested with SgeI, 1.4% agarose, 6 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

DNA methylated by Dcm or CpG methyltransferases will be a substrate for SgeI. Greater than 3-fold overdigestion with SgeI may result in incomplete cleavage. At least two copies of SgeI recognition sequence are required for an efficient cleavage. Amount of the enzyme required for complete digestion of methylated DNA depends on the number of SgeI recognition sites. DNA cleavage products generated by target site cleavage facilitate the nonspecific cleavage by SgeI.

Therefore, optimization of the enzyme amount is recommended for DNA cleavage. pBR322 DNA isolated from E. coli dcm+ strain (#SD0041) can be used as a DNA cleavage efficiency control. SgeI cleaves all six dcm methylated targets on pBR322 DNA.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer SgeI: 10 mM Tris-HCl (pH 8.0 at 37°C), 5 mM MgCl2, 100 mM KCl, 0.02% Triton X-100, and 0.1 mg/mL BSA
  • Incubate at 37°C
Digestion of Agarose-embedded DNAMinimum 3 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded pBR322 DNA isolated from E. coli dcm+ strain DNA in 16 hours at 37°C.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferSgeI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM NaCl, 1 mM DTT, 1 mM EDTA, 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
SgeI Buffer (Unique) 37°C 0-20 0-20 0-20 NR NR NR NR

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm (CCWGG)Cm5CWGG Cleaves only methylated DNA
CpG 5'...m5C GNG...3'
3'... Gm5CNC...5'		 Cleaves only methylated DNA
CpG 5'...m5C Gm5C G...3'
3'... Gm5CGm5C...5'		 Cleaves only methylated DNA