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. 2000 Sep;20(18):6806–6815. doi: 10.1128/mcb.20.18.6806-6815.2000

FIG. 4.

FIG. 4

Effects of salt, primer, and nucleotide concentration on the coupled cleavage and extension reactions mediated by yeast telomerase. (A) Polymerization assays were carried out using OXYT1 as the DNA primer and DEAE column fractions as the source of telomerase. For reactions 1 to 3, the DEAE fraction was first desalted using Centricon-30. Sodium acetate was then added to the following final concentrations: lane 1, 0 mM; lane 2, 150 mM; lane 3, 300 mM. For reactions 4 to 7, the following concentrations of OXYT1 primer were used: lane 4, 3 μM; lane 5, 6 μM; lane 6, 12 μM; lane 7, 24 μM. (B) Polymerization assays were carried out using OXYT1 as the DNA primer and DEAE column fractions as source of telomerase. In addition to 0.2 μM labeled dGTP (3,000 Ci/mmol; NEN), unlabeled dGTP was added to the following concentrations: lane 1, 0 μM; lane 2, 0.5 μM; lane 3, 1.0 μM; lanes 4 and 5, 2.0 μM. Lane 5 represents a longer exposure than lane 4.