DNA Polimerase I - Large (KLENOW) Fragment
Possesses:
-
3’ ? 5’ exonuclease activity
-
5'-3' polymerase activity
Reaction buffer (1X E.coli Polymerase
I/Klenow buffer)
-
10 mM Tris-HCl (pH 7.5)
-
5 mM MgCl2
-
7.5 mM dithiothreitol
Supplement with dNTP’s
Klenow is also 50% active in all
four standard NEB buffers.
Heat inactivation by incubating
at 75°C for 20 minutes.
Fill in conditions
-
33 mM each dNTP
-
1 unit of Klenow per mg DNA
-
incubate at 25°C for 15 minutes