What is the effect of high temperature during DNA extraction?

Incubation at temperatures above ~45°C during DNA extraction can lead to the generation of single-stranded DNA. This can have the same effect as protein contamination, leading to increased risk of sequencing problems, including an inability to deliver CNV and SV results and higher small variant error rates. Some protocols recommend incubations with Proteinase K at higher temperatures. In this case, it is important to limit the amount of time the DNA sample spends at the higher temperature, ideally to a maximum of a couple of hours.


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