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Determination of the RNA primer initiation site on the TTA template for S. aureus primase. (A) Schematics of the predicted RNA primer product (italics) in either the presence or absence of rCTP (bold) on the ssDNA template. (B) Denaturing HPLC analysis of RNA primer synthesis in the presence of 0, 25, 50, 100, 200, and 400 μM rCTP. The reactions were performed using 2 μM ssDNA template, 2 μM primase, and 400 μM rNTPs during a 1 -h incubation at 30°C. Abs, absorbance at 260 nm.
