Recently Viewed

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

MboII

5'...G A A G A (N)8...3'
3'...C T T C T (N)7...5'

Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available B buffer for 100% activity Optimal incubation at 37°C Star activity Cleavage blocked or impaired by overlapping Dam methylation Thermal inactivation at 65°C in 20 min Recombinant enzyme LO certified

The MboII restriction enzyme recognizes GAAGA(8/7)^ sites and cuts best at 37°C in B buffer
  
Loading...
Lambda DNA (dam-) digested with MboII

Lambda DNA (dam-) digested with MboII, 1.4% agarose, 130 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 lambda DNA (dam) (#SD0021). MboII is blocked by overlappingdam methylation. To avoid dam methylation, use a dam, dcm- strain. Greater than 15-fold overdigestion with MboII may result in star activity. MboII produces DNA fragments that have a single-base 3 extension, which are more difficult to ligate than blunt-ended fragments.

MboII 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 B: 10mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, and 0.1mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferMboII is supplied in: 10mM Tris-HCl (pH 7.5 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/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
B (Blue) 37°C 100 50-100 20-50 0-20 50-100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dam (GATC) 5'...GAAGm6A TC...3'
3'...CTTC Tm6AG...5'
Blocked
CpG 5'...m5C GAAGA...3'
3'... Gm5CTTCT...5'
No effect
EcoBI (TGA(N)8TGCT) 5'...TGm6AAGA(N)5 TGCT...3'
3'...AC TTCT(N)5m6ACGA...5'
No effect

Cleavage efficiency close to the termini of PCR fragments

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

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
130 11 11 11 7 8
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
9 9 8 8 15 11