Effect of N3-Methyladenine and an Isosteric Stable Analogue on DNA Polymerization
Figure 5
Insertion opposite 3-mA by T7 Sequenase. The template strand was treated with Me-lex as described in Figure 1. A -Cy5-primer (Table 1, P25) was added that overlaps with the two A’s (A24, A25) that are not methylated by Me-lex. T7 Sequenase was added and the reactions initiated by the addition of 5, 50 or 500 M concentration of the individual dNTPs at 37°C. The insertion of T is inhibited in the 2.5 mM Me-lex treated DNA versus the untreated DNA (0 Me-lex). There is no indication that G, T or C is inserted opposite the lesion.
