After running Nextera Flex my product size is way too big, why?

08 October 2020 1 7K Report

Hiya!

I'm currently running my samples through a Nextera Flex library prep in order to sequence them. My samples have been whole genome amplified using the Sigma WGA4 kit, bead-cleaned, and then into the Nextera protocol. Before Nextera, sample sizes are averaging at a quite expected 155bp, but the tapestation following the Nextera have them averaging at about 1500bp, just by the upper ladder (not for negative controls). Attaching examples of 1 sample.

Has anyone experienced something similar? Any ideas what may have caused this?

Thank you for your inputs and advice in advance!

Joar Franzén

Found I had accidentally run our PCR default at 25 cycles instead of the 8 that I was supposed to set it to... Probably explains it.

Thanks anyhow!

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