ACC65 I (2000 U)

Descrizione

G/GTACC
CCATG/G

Source:
An E. coli strain that carries the cloned Acc65I gene from Acinetobacter calcoaceticus 65 (S.K. Degtyarev).

Reagents Supplied:
NEBuffer 3 (10X)
BSA (100X)


Enzyme Properties


Activity in NEBuffers:
NEBuffer 1:10%
NEBuffer 2:75%
NEBuffer 3:100%
NEBuffer 4:25%

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: Blocked by some combinations of overlapping
CpG methylation: Blocked by some combinations of overlapping

Heat Inactivation:
65°C for 20 minutes

Survival in a Reaction:
(+) Intermediate activity. Suitable for extended digestion, but < 8 hours.
More information about: Extended Digests with Restriction Enzymes


Reaction & Storage Conditions


Reaction Conditions:
1X NEBuffer 3
Supplemented with 100 ?g/ml Bovine Serum Albumin
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 pBC4 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
10,000 units/ml

Unit Assay Substrate:
pBC4 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:
Acc65I is an neoschizomer of KpnI.