How critical is the 65°C denaturation step in cDNA synthesis?

07 November 2023 0 6K Report

Hi All,

I was synthesizing cDNAs and after denaturation at 65°C for 5 min, I was late for immediately placing the tubes on ice. The temperature decreased to 50°C when I was transferring tubes on ice. Does anybody know if this gradual decrease in the temperature (instead of immediate cooling on ice) could be a reason for failure in the cDNA synthesis?

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