Equipamentos para Laboratórios | Equipamento para Laboratório

Recognition Site:

Source:
Arthrobacter citreus (C. Polisson)

Reagents Supplied:
NEBuffer 3

Enzyme Properties
Activity in NEBuffers:
NEBuffer 1:        25%
NEBuffer 2:        50%
NEBuffer 3:        100%
NEBuffer 4:        50%

When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

Methylation Sensitivity:
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG methylation: Blocked
More information about: Methylation Sensitivity

Heat Inactivation:
65°C for 20 minutes

Survival in a Reaction:
(–) Not recommended for digest over 1 hour.
More information about: Extended Digests with Restriction Enzymes

Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 3
Incubate at 37°C.

1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
10,000 units/ml

Unit Assay Substrate:
λ DNA

Storage Conditions:
10 mM Tris-HCl
100 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Storage Temperature:
-20°C

Diluent Compatibility:
Diluent A

Notes
General notes:

1. AciI has a non-palindromic recognition site. As a result, when DNA is cut with AciI and then ligated, only 50% of these ligated sites regenerate an AciI site.

Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.

Ligation and Re-cutting:
After a 10-fold overdigestion with AciI, > 95% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, approximately 50% can be recut with AciI.

The remaining ligation products form HpaII/MspI sites.

16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 40 units of AciI incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

Exonuclease Activity:
Incubation of a 50 μl reaction containing 50 units of AciI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity.

Reagents Sold Separately
NEBuffer 3

AciI FAQ

Q1: Why can only 50% of AciI cut and ligated fragments be recleaved by AciI?
A1: AciI has a non-palindromic recognition site. As a result, when DNA is cut with AciI and then ligated, only 50% of these ligated sites regenerate an AciI site.