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Figure 4
Cell cycle-related changes in G-overhang length. (A) Primer extension products obtained using cells synchronized by centrifugal elutriation: lanes 1–3, DNA isolated from three different batches of S phase cells; lanes 4–6, DNA isolated from a single batch of elutriated cells after 30 min growth (lane 4), 60 min growth (lane 5) and 90 min growth (lane 6); lane 7, DNA from unsynchronized exponentially growing cells; lane 8, DNA from starved cells; lanes 9–12, longer exposure of lanes 4–7. Positions of markers are shown on the left. The overhang lengths corresponding to specific reaction products are indicated on the right. (B) Distribution of G-overhangs throughout the cell cycle. G1, S and G2, the reaction products in lanes 4–6 of (A) were quantified by PhosphorImager analysis and the total amount of reaction product in each pair of bands (i.e. the 14 and 15 nt products, the 20 and 21 nt products, etc.) was normalized to the total amount of product in that lane: G1, cells in G1 and early S phase (50% BrdU staining); S phase, 95% BrdU staining; G2, cells in G2 and late S (40% BrdU staining); Unav, Stav, Sav, the reaction products were quantified from four different batches of unsynchronized growing cells, starved cells and cells synchronized in S phase.
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