Removal of single stranded extensions using Mung
Bean Nuclease
Mung Bean endonuclease degrades
single-stranded extensions from the ends of DNA and RNA molecules,
leaving blunt, ligatable ends.
1. DNA (0.1 ug/ul) should be suspended in either 1xMung
bean buffer:
-
50 mM sodium acetate pH 5.0
-
30 mM NaCl
-
1 mM ZnSO4
or in NEB buffers 1, 2 or 4. If
using NEBuffer 1 2 or 4, add ZnSO4 to final conc. Of 1mM
Reaction volume should be 50-100µl
2. Add 1u of enzyme
3. Incubate at 30deg. 30min.
4. Inactivate the enzyme by phenol/chloroform extraction
5. Recover DNA by ethanol precipitation