The algorithm of estimation of the Km values for primers of various structure and length in the polymerization reaction catalyzed by Klenow fragment of DNA polymerase I from E.coli

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Abstract
DNA synthesis at primers d(pT) n , d(pA) n , d(pC) n , and d(pG) n in the presence of corresponding complementary templates and at hetero‐oligoprimers complementary to MB phage DNA was investigated. The values of both ‐log K m and log V max increased linearly if homo‐oligoprimers contained less than 10 nucleotides. The lengthening of d(pT) n and d(pA) n primers by one mononucleotide unit (n = 1–10) resulted in the 1.82‐fold decrease of the K m values. The incremental decreases of K m for d(pC) n and d(pG) n were equal to about 2.46. The enhancement of the homo‐ and heterooligonucleotide primers' affinity to the enzyme due to one Watson‐Crick hydrogen bond between complementary template and primer is about 1.35 times. This allows to calculate the K m values for primers of various structure and length up to 10 units. The objective laws of the K m and V max values changes for primers containing more than 10 nucleotides were analyzed.