(Neisseria mucosa heidelbergensis)
5’-G C T A G C-3’
3’-C G A T C G-5’
Recombinant. Purified from E.coli strain carrying the NheI gene from Neisseria nucosa heidelbergensis.
Reaction Temperature: 37°C
Prototype: NheI
Inactivation Temperature (20 min): 65°C
Reaction Buffer:
1x NheI Buffer: 20 mM Tris-HCl (pH 7.4 at 25°C), 5 mM MgCl2, 50 mM KCl, 100 µg/ml bovine serum albumin.
Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of Ad-2 DNA in 1 hr in a total reaction volume of 20 µl.
Notes:
- due to star activity it is not recommended to incubate over 4 hours or to use more than 100 units per reaction;
- salt concentration above 50 mM inhibits NheI activity.
Assay Conditions: 20 mM Tris-HCl (pH 7.4 at 25°C), 50 mM KCl, 5 mM MgCl2, 100 µg/ml bovine serum albumin and 1 µg of Ad-2 DNA. Incubation is at 37°C for 1 hr in a reaction volume of 20 µl.
Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 25°C), 250 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.1% (v/v) Triton™X-100, 500 µg/ml bovine serum albumin and 50% (v/v) glycerol.
Storage Conditions: Store at –20°C.
Quality Control:
All preparations are assayed for contaminating endonuclease, 3'-exonuclease, and nonspecific single- and