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Anal Biochem. 1990 Dec;191(2):396-400.

Purification of Thermus aquaticus DNA polymerase expressed in Escherichia coli.

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1
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.

Abstract

DNA polymerase from Thermus aquaticus has become a common reagent in molecular biology because of its utility in DNA amplification and DNA sequencing protocols. A simplified method is described here for isolating the recombinant Taq enzyme after overproduction in Escherichia coli. Purification requires 8 to 10 h and entails heat treating and clearing the E. coli lysate, followed by precipitation of the enzyme with polyethyleneimine and elution from Bio Rex 70 ion exchange resin in a single salt step. The resulting enzyme preparation contains a single, nearly homogeneous protein consistent with the previously established size of the Taq DNA polymerase in a yield of 40-50 mg of protein per liter of cell culture.

PMID:
2085185
[Indexed for MEDLINE]
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