MboII
Detailní popis
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.
| Conditions for 100% Activity |
|
| Double Digestion | Perform double digestion using DoubleDigest. |
| Hazardous | No |
| Isoschizomers | Search for commercial isoschizomers using REsearch. |
| Storage Buffer | MboII 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 |
| Storage Condition | -20 C |
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
- Dam: may overlap - blocked.
- Dcm: never overlaps - no effect.
- CpG: may overlap - no effect.
- EcoKI: never overlaps - no effect.
- EcoBI: may overlap - no effect.
| 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 |
Hodnocení produktu
Produkt zatím nikdo nehodnotil, buďte první!