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BstXI

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

FastDigest enzyme available O buffer for 100% activity Optimal incubation at 55°C Star activity Cleavage blocked or impaired by overlapping Dcm methylation Thermal inactivation at 80°C in 20 min Genome qualified LO certified

The BstXI restriction enzyme recognizes CCANNNNN^NTGG sites and cuts best at 55°C in O buffer
  
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Lambda DNA digested with BstXI

Lambda DNA digested with BstXI, 0.7% agarose, 13 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

Incubation at 37°C results in 50% activity. Greater than 15-fold overdigestion with BstXI may result in star activity. BstXI cleavage is impaired 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
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 55°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.
Storage BufferBstXI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 50 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
O (Orange) 55°C 20-50 100 100 50-100 50-100 100 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...Cm5CA GG(N)4TGG...3'
3'...G GTm5CC(N)4ACC...5'		 No effect
Dcm (CCWGG) 5'...Cm5CA GGNNCm5CT GG...3'
3'...G GTm5CCNNG GAm5CC...5'		 Blocked

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
13 3 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 1 1 0 0

 

MSDS & Certificates

pdf   BstXI - MSDS
pdf   MSDS (JP) - Restriction Enzymes