Klenow Fragment (3′→5′exo-)
Klenow fragment (3' to 5' exo-) is proteolytic product of E. coli DNA Polymerase I which retains polymerization and 5' to 3' exonuclease activity, but has lost 3' to 5' exonuclease activity with D355A and E357A mutation.
A recombinant E.coli strain that carries polA gene fragment with D355A and E357A mutation.
• DNA sequencing by the Sanger dideoxy method
• Second strand synthesis of cDNA;
• Synthesis of random primers
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 37°C.
1×Klenow fragment (3' to 5' exo-) Reaction Buffer: [10 mM Tris-HCl (pH 7.9 at 25°C)，50 mM NaCl，10 mM MgCl2，1 mM DTT]; add dNTPs (not supplied with enzyme)，incubate at 37°C.
Store at -20°C