BveI (BspMI)
Kód produktu: ER1741
Kód výrobce: ER1741
Kód dodavatele: {C5AC70B1-0845-45C2-90A6-93443D0D7302}
Výrobce:
Life Technologies Czech Republic s.r.o.
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
Notes
- At least two copies of BveI recognition site are required for efficient cleavage. Inclusion of 0.5 µM oligonucleotide with the BveI recognition sequence in the reaction mixture significantly improves cleavage of plasmid DNAs, especially of those with a single BveI site. Still, complete cleavage of some substrates with BveI is difficult to achieve.
- BveI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase.
- Low salt, high glycerol ( > 5%) concentrations, pH > 8.0, or a large excess of enzyme may result in star activity.
- BveI 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 | BveI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 150 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/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 |
||||
| O (Orange) +oligo | 37°C | 0-20 (+oligo) | 20-50 (+oligo) | 100 (+oligo) | 20-50 (+oligo) | 50-100 (+oligo) | 100 (+oligo) | 1X or 2X (+oligo) |
Methylation Effects
- Dam: never overlaps - no effect.
- Dcm: never overlaps - no effect.
- CpG: may overlap - cleavage impaired.
- EcoKI: may overlap - effect not determined.
- EcoBI: may overlap - effect not determined.
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
| CpG | 5'...ACCTGm5C G...3'3'... TGGAC Gm5C...5' |
Impaired |
Cleavage efficiency close to the termini of PCR fragments
| bp from the recognition site to fragment end | ||||
|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 |
| 0-20 | 50-100 | |||
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 41 | 3 | 3 | 1 | 1 | 0 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(±) | pBluescriptIISK(±) | pACYC177 | pACYC184 |
| 1 | 0 | 0 | 0 | 0 | 1 |
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