Eco31I (BsaI)
Kód produktu: ER0291
Kód výrobce: ER0291
Kód dodavatele: {404B22CD-1F16-4EB9-A2B2-1B9EF140099B}
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
Note
- Assayed using lambda DNA (dcm-) (#SD0021) as one of the two Eco31I recognition sites in lambda DNA is difficult to cleave. Eco31I cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.
- Low salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.
- Eco31I cleaves downstream of its recognition site and can generate any desired 4 base 5'-overhangs. This feature is useful for direct PCR product cloning.
For methylation sensitivity refer to product specifications.
| Conditions for 100% Activity |
|
| Digestion of agarose-embedded DNA | Minimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours. |
| Double Digestion | Perform double digestion using DoubleDigest. |
| Hazardous | No |
| Isoschizomers | Search for commercial isoschizomers using REsearch. |
| Storage Buffer | Eco31I is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 200 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 |
||||
| G (Green) | 37°C | 50-100 | 100 | 0-20 | 0-20 | 50-100 | 20-50 | 1X or 2X |
Methylation Effects
- Dam: never overlaps - no effect.
- Dcm: may overlap - cleavage impaired.
- CpG: may overlap - cleavage impaired.
- EcoKI: never overlaps - no effect.
- EcoBI: may overlap - effect not determined.
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
Dcm (CCWGG) |
5'...Cm5CW GGTCTC...3'3'... G GWm5CCAGAG...5' |
Impaired |
| CpG | 5'...GGTCTm5C G...3'3'... CCAGA Gm5C...5' |
No effect |
| CpG | 5'...m5C GGTCTC...3'3'... Gm5CCAGAG...5' |
Impaired |
Cleavage efficiency close to the termini of PCR fragments
| bp from the recognition site to fragment end | ||||
|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 |
| 20-50 | 50-100 | |||
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 2 | 0 | 0 | 1 | 1 | 1 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
| 1 | 1 | 1 | 1 | 1 | 0 |
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