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. 1988 Mar-Apr;5(2):32-9.
doi: 10.1016/0735-0651(88)90024-6.

Optimal conditions for supercoil DNA sequencing with the Escherichia coli DNA polymerase I large fragment

H M Lim  1 , J J Pène
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Optimal conditions for supercoil DNA sequencing with the Escherichia coli DNA polymerase I large fragment

H M Lim et al. Gene Anal Tech. 1988 Mar-Apr.

Abstract

Sequencing ladders produced from supercoiled DNA templates with the Escherichia coli DNA polymerase Klenow fragment are often unreadable because of a high background and misincorporated nucleotides. This study showed that contaminating RNA molecules can interfere with template:primer hybridization. Procedures are provided for the purification of template DNA and stringent conditions for primer-template hybridization that overcome these problems.

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