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Klenow Fragment


Klenow fragment
A proteolytic fragment of DNA polymerase I that contains the DNA polymerase activity and the 3' to 5' proofreading activity but lacks the 5' to 3' exonuclease activity. This enzyme is widely used for DNA sequencing.
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Klenow Fragment: The large fragment of E. coli DNA polymerase I which lacks 5' -> 3' exonuclease activity. Very useful for sequencing reactions, which proceed in a 5' -> 3' fashion (addition of nucleotides to templated free 3' ends of primers).
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In contrast, two other polymerases used for DNA sequencing, Klenow fragment of E. coli DNA polymerase I and Taq DNA polymerase, discriminate against dideoxynucleotides up to one thousand fold. This results in marked variability in band intensities.
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