Klenow Fragment 3'→5' exo-

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Cat. NO. AB-RK20526 (datasheet)

Klenow Fragment (3'→ 5' exo-) is an N-terminal truncation of DNA Polymerase I which retains polymerase activity, but has lost the 5'→ 3' exonuclease activity and has mutations (D355A, E357A) which abolish the 3'→ 5' exonuclease activity. Klenow Fragment (3'→ 5' exo-) is isolated from a recombinant source. It generates probes using random primers and shows moderate strand displacement activity. It can be used in random primer labeling, DNA sequencing by the Sanger dideoxy method, second strand cDNA synthesis and second strand synthesis in mutagenesis protocols.

Featured Specification
Product Source An E.coli strain containing a plasmid with a fragment of the E.coli polA (D355A, E357A) gene starting at codon 324.
Applications A-tailing, Polymerases for DNA Manipulation, DNA Sequencing , RT-PCR & cDNA Synthesis , PCR
Unit Definition One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid insoluble material in 30 minutes at 37°C.
Storage Conditions 25 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4 @25℃
Storage Temperature -20°C
Reaction Conditions 1x ABuffer B, Incubate at 37°C
Reaction Buffer or Mix 1X ABuffer B:
10 mM Tris-HC, 50 mM NaCl, 10 mM MgCl2, 1 mM DTT, pH7.9 @ 25°C
Heat Inactivation 75°C for 20 min
5' - 3' Exonuclease No
3' - 5' Exonuclease No
Strand Displacement +++
Protein M.W. Theoretical 68000 daltons
Error Rate ~ 100 x 10-6 bases