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SgrDI

5'...C G↓T C G A C G...3'
3'...G C A G C T↑G C...5'

R buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by CpG methylation Thermal inactivation at 65°C in 20 min Genome qualified Recombinant enzyme LO certified

The SgrDI restriction enzyme recognizes CG^TCGACG sites and cuts best at 37°C in R buffer
  
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Lambda DNA digested with SgrDI

Lambda DNA digested with SgrDI, 0.7% agarose, 1 cleavage site

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

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsSalI, Eco88I, SmiI, XhoI.
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
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteLow salt concentration or large excess of the enzyme may result in star activity.
Storage BufferSgrDI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 mM KCl, 1 mM DTT, 0.1 mM 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
R (Red) 37°C 0-20 0-20 0-20 100 NR 100 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpGCGTCGACG Blocked

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
1 0 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 0

New sites generated by ligation

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

Recognition SequenceSecond EnzymeEnzymes recognizing newly generated recognition sequence
CG^TCGACG
  • AbsI (CC^TCGAGG)
  • Eco88I (AvaI)/FastDigest AvaI (Eco88I)* (C^TCGAG)
  • PspXI* (VC^TCGAGC)
  • PspXI* (VC^TCGAGG)
  • PspXI* (VC^TCGAGT)
  • SmoI (SmlI)* (C^TCGAG)
  • XhoI/FastDigest XhoI (C^TCGAG)
  • Hpy99I
  • TaqI/FastDigest TaqI
  • SalI/FastDigest SalI (G^TCGAC)
  • HincII (HindII)/FastDigest HincII
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • Hpy99I
  • SalI/FastDigest SalI
  • TaqI/FastDigest TaqI
  • XmiI (AccI)/FastDigest AccI (XmiI)

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

Recognition SequenceNewly generated sequence after reactionEnzymes that cleave the newly generated sequence
CG^TCGACGCGTCGATCGACG
  • Bsh1285I (BsiEI)/FastDigest BsiEI (Bsh1285I)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • [Hpy99I]
  • MboI/FastDigest MboI
  • PvuI/FastDigest PvuI
  • [TaqI/FastDigest TaqI]