APex Phosphatase can be added directly to most restriction
enzyme buffers and is highly active over a broad range of temperatures
(up to 50°C), pH (from 5-12), and salt conditions (e.g., up to 1
M Na+, NH4+, K+, Cl-,
or OAc-), and in the presence of 10% Triton® X-100. To
remove the 5´ phosphate from DNA, RNA, or a nucleotide, simply
incubate the substrate with APex Phosphatase at 37°C for 10 minutes.
The same reaction conditions may be used for all types of DNA ends, including
difficult 5´-recessed ends.
Unit Definition: One microliter of APex Heat-Labile Alkaline
Phosphatase dephosphorylates 1 µg of pUC19 vector DNA digested
with Hind III (5´ protruding), Hinc II (blunt), or Pst I
(5´recessed) in 10 minutes at 37°C.
Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5),
0.1 mM zinc acetate, 10 mM MgCl2, 0.1 M NaCl, and 0.1% Triton® X-100.
10X Reaction Buffer: 330 mM Tris-acetate (pH 7.8), 660 mM potassium
acetate, 100 mM magnesium acetate, and 5.0 mM DTT.
Quality Control: Function-tested to meet EPICENTRE's Cloning
Quality Standard of greater than 95% inhibition of self-ligation and
checked by transformation into E. coli. Free of detectable exo-
and endonuclease and RNase activities. |
| 5´-(4-base) overhang: |
Hind III, EcoR I |
| 5´-(3-base) overhang: |
EcoO109 I |
| 5´-(2-base) overhang: |
Nde I |
| 3´-(4-base) overhang: |
Sph I, Sac I, Pst I |
| 3´-(3-base) overhang: |
AlwN I |
| 3´-(1-base) overhang: |
Ahd I |
|
