Aju I
5'...↓ 7(N) G A A (N)7 T T G G (N)11↓...3'
3'...↓12(N) C T T (N)7 A A C C (N)6 ↓...5'
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
Complete cleavage of some substrates with AjuI is difficult to achieve. AjuI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase. AjuI produces double-strand cuts on both sides of the interrupted recognition site. In certain sequence contexts, the cleavage position may be shifted by one base pair. However, the cleavage position indicated above will predominate.
| 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. |
| Note |
|
| Storage Buffer | AjuI is supplied in 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 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 |
||||
| R (Red) + SAM | 37°C | 0-20 (+SAM) | 50-100 (+SAM) | 20-50 (+SAM) | 100 (+SAM) | 50-100 (+SAM) | 50-100 (+SAM) | 1X or 2X (+SAM) |
Methylation Effects
- Dam: never overlaps - no effect
- Dcm: never overlaps - no effect
- CpG: may overlap - no effect
- EcoKI: never overlaps - no effect
- EcoBI: never overlaps - no effect
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
| CpG | 5'...m5C GAA(N)7TTGG...3'3'... Gm5CTT(N)7AACC...5' |
No effect |
Number of recognition sites in DNA molecules
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 3 | 1 | 0 | 0 | 0 | 0 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(±) | pBluescriptIISK(±) | pACYC177 | pACYC184 |
| 0 | 0 | 0 | 0 | 1 | 0 |
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