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AAT Bioquest

What causes over-fragmented DNA during sequencing library construction?

Posted February 17, 2022


Answer

High temperatures and compromised sample integrity are two of the main causes of over-fragmented DNA during sequencing library construction. During sequencing library construction, the reaction should be kept on ice for best results. If the reaction is left at room temperature instead, over-fragmentation can occur. Samples with compromised integrity have a higher tendency to result in over-fragmented DNA.

Additional resources

Library construction for next-generation sequencing: Overviews and challenges

MagaDye™ 4 Color Sanger Sequencing Terminator Kit