FaqI (BsmFI)
Kód produktu: ER1811
Kód výrobce: ER1811
Kód dodavatele: {8C31FDFD-1991-43E8-BC0C-FF3669EB4369}
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
- FaqI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05mM S-adenosylmethionine gives more than a 2-fold increase in FaqI activity. Still, complete cleavage of some substrates is difficult to achieve. FaqI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase.
- FaqI 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 | FaqI 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. |
| 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 |
||||
| Tango | 37°C | 20-50 (+SAM) | 20-50 (+SAM) | 0-20 (+SAM) | 0-20 (+SAM) | 100 (+SAM) | 50-100 (+SAM) | 1X (+SAM) |
Methylation Effects
- Dam: never overlaps - no effect.
- Dcm: may overlap - no effect.
- CpG: may overlap - blocked.
- EcoKI: never overlaps - no effect.
- EcoBI: never overlaps - no effect.
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
Dcm (CCWGG) |
5'...Cm5CW GGGAC...3'3'... G GWm5CCCTG...5' |
No effect |
| CpG | 5'...GGGAm5C G...3'3'... CCCT Gm5C...5' |
Blocked |
| CpG | 5'...m5C GGGAC...3'3'... Gm5CCCTG...5' |
Blocked |
Cleavage efficiency close to the termini of PCR fragments
| bp from the recognition site to fragment end | ||||
|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 |
| 0-20 | 50-100 | |||
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 38 | 2 | 2 | 4 | 0 | 0 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
| 0 | 0 | 0 | 0 | 1 | 7 |
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