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OPTIZYME™ AloI, Fisher BioReagents™
Brand: Fisher BioReagents BP8075-1
This item is not returnable.
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Description
5'...^ 7(N) G A A C (N)6 T C C (N)12-13^...3'
3'...^12-13(N) C T T G (N)6 A G G (N)7 ^...5'
Supplied with: 10X OPTIZYME Buffer 5
Conditions for 100% Activity:
- 1X OPTIZYME Buffer 5: 10mM Tris-HCl (pH 8.5 at 37°C), 10mM MgCl2, 100mM KCl and 0.1mg/mL BSA
- Incubate at 30°C
- Buffer 1: 0 - 20%
- Buffer 2: 0 - 20%
- Buffer 3: 0 - 20%
- Buffer 4: 20 - 50%
- Buffer 5: 100%
- 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
- Incubation at 37°C results in 20% activity.
- AloI produces double-strand cuts on both sides from the interrupted recognition site. Its unique feature is a degenerate cleavage point on the 3 side of the recognition sequence (12 or 13 nucleotides away).
- The presence of S-adenosylmethionine in the reaction mixture results in incomplete cleavage with AloI.
- Greater than 10-fold overdigestion with AloI may result in star activity.
- AloI 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.
Dam: May overlap - effect not determined
- 5'...GAAC(N)4Gm6A TCC...3'
- 5'...GAAC(N)6TCm5CW GG...3'
- 5'...m5C GAAC(N)6TCC...3'
- 5'...GAAC(N)6TCm5C G...3'
- 5'...GAAm5C G(N)5TCC...3'
Specifications
1 to 3 U/μL | |
30°C | |
>95% of DNA fragments can be ligated and re-cut after 50-fold over-digestion with AloI | |
Keep container tightly closed | |
(7/12)GAAC(N)6TCC(12/7) | |
Liquid |
25 to 50% Water, >50% Glycerin | |
7.4 | |
10mM Tris HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA, 50% Glycerol | |
100 U | |
AloI |