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. 2005 Jan 26;33(2):525–535. doi: 10.1093/nar/gki178

Figure 8.

Figure 8

HPLC analysis of products of 5′-CCCCCCCCCCCCA-3′–5′-GGGGTGGGGGGGA-3′ extension. Polymerase extension assay was performed as described in Materials and Methods with 5 μM 5′-CCCCCCCCCCCCA-3′–5′-GGGGTGGGGGGGA-3′ and 10 μg/ml of Klenow exo− at 37°C. The reaction was started by addition of the enzyme and terminated by addition of 10 mM EDTA. Aliquots of 50 μl of sample were withdrawn from the assay mixture before (black curve) and 10 (red curve), and 20 (blue curve) min after the reaction had been started. Oligomers were separated from dGTP and dCTP with TSKgel G-3000 SWXL HPLC column (7.8 × 300 mm) and loaded in 0.1 M KOH onto TSKgel DNA-NPR column (4.6 × 75 mm). Elution was performed using a 20 min linear gradient between 0 and 1 M KCl in 0.1 M KOH at a flow rate of 0.6 ml/min.