Aar I
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
For methylation sensitivity refer to product specifications.
For cleavage with AarI at least two copies of its recognition sequence are required. Inclusion of 0.5 µM oligonucleotide with the AarI recognition sequence in the reaction mixture significantly improves cleavage of DNAs, especially of those with a single AarI site.
Still, a complete cleavage of some substrates with AarI is difficult to achieve. Greater than 10-fold overdigestion with AarI may result in star activity. AarI 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.
| Conditions for 100% Activity |
|
| Double Digestion | Perform double digestion using DoubleDigest. |
| Hazardous | No |
| Isoschizomers | Search for commercial isoschizomers using REsearch. |
| Storage Buffer | AarI is supplied in: 10 mM potassium phosphate (pH7nbsp;7.4 at 25°C), 100 mM KCl, 1 mM EDTA, 1 mM DTT, 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 |
||||
| AarI +oligo Buffer (Unique) | 37°C | NR (+oligo) | NR (+oligo) | 0-20 (+oligo) | 0-20 (+oligo) | NR (+oligo) | 50-100 (+oligo) | 2X (+oligo) |
Methylation Effects
- Dam: never overlaps - no effect
- Dcm: never overlaps - no effect
- CpG: may overlap - cleavage impaired
- EcoKI: never overlaps - no effect
- EcoBI: may overlap - effect not determined
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
| CpG | 5'...CACCTGm5C G...3'3'... GTGGAC 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 |
| 20-50 | 50-100 | |||
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 12 | 0 | 0 | 0 | 0 | 0 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(±) | pBluescriptIISK(±) | pACYC177 | pACYC184 |
| 0 | 0 | 0 | 0 | 0 | 0 |
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