Comparison Of Constant- And PulsedField Electrophoresis Technologies For Analysis Of High Molecular Weight And Large DNA Fragments

By Timothy Butler, Agilent Technologies

Understanding DNA fragment size is essential for confident decision‑making in NGS workflows, especially when working with high molecular weight (HMW) samples. Explore how different electrophoresis technologies handle large DNA fragments and why choosing the right method can make or break long‑read sequencing success. Compare constant‑field and pulsed‑field separation, highlighting how compression in constant‑field methods can distort sizing above ~10 kb and lead to inaccurate assessments of sheared or genomic DNA. By examining real examples across 10–20 kb smears, see where each approach excels and where common QC challenges arise.

For anyone preparing long‑read libraries or evaluating HMW DNA integrity, gain insight into selecting the most accurate sizing method.