DNA fragments of varying AT content were custom-ordered from Integrated DNA Technologies and were amplified by high-fidelity PCR. Briefly, the 20-μl reaction mixture contained 1 ng of sample DNA, forward and reverse primers (10 μM), 10 mM deoxynucleoside triphosphate (dNTPs) and 20 U of Phusion High-Fidelity Polymerase (New England Biolabs, no. M0530S) with the provided 5× Phusion High-Fidelity or GC buffer. PCR conditions were as follows: initial denaturation 98°C for 30 s, denaturation at 98°C for 10 s, annealing at 65°C for 20 s, extension at 72°C for 30 s, and final extension at 72°C for 5 min, for a total of 20 cycles. The products were purified using Agencourt AMPure XP Beads (Beckman Coulter, no. 63881), at 0.6× beads-to-product ratio.

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