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This study examined the bypass of γ-OH-PdG by Sulfolobus solfataricus Dpo4, the prototypic Y-family DNA polymerase, using templates that contained the adduct in either the 5′-CXG-3′ or the 5′-TXG-3′ sequence context.
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To further explore the effects of local sequence context on the kinetic signatures of 5mC and 5caC, we used a synthetic SMRTbell template that contained a modified base in a 5'-CG-3' sequence context, surrounded by two random bases on each side.
Templates were prepared using forward primers that contained the T7 RNA polymerase promoter sequence (CCAAGCTTCTAATACGACTCACTATAGGGAGA [T7]).
The DNA templates were generated by PCR from cDNA clone LD 40196 (γ-tubulin 23C), using PCR primers that contained the T7 RNA polymerase minimal promoter sequence.
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