What are the main factors causing the size instability of enzymatic digestion?

• Sample quality difference: Whether the samples are degraded. For degraded samples, the enzymatic digestion time can be shortened in a gradient of 3-5 min according to the recommended time in the user manual.
• Sample solvent: Solvent contains EDTA or pH is too high (pH greater than 8.0, such as pH 8.5). When the sample solvent contains EDTA, if it is not purified, Enhancer Buffer is not added, or the solvent pH is too high, it will lead to insufficient fragmentation.

A. 50 ng blood gDNA samples containing EDTA (0-0.8 mM) were used for library preparation. The higher the EDTA concentration, the larger the enzymatic digestion fragment.