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Updated: Jun 8, 2026

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
Published on: July 3, 2016
Denaturing HPLC for mutation screening
Mike Mitchell1, Jacqueline Cutler
1Molecular Genetics, Centre for Haemostasis & Thrombosis, St. Thomas' Hospital, Guy's & St Thomas' NHS Foundation Trust, London, UK.
Abstract:
Denaturing High-Performance Liquid Chromatography (dHPLC) is probably the most versatile and one of the most widely used mutation screening technologies. It benefits from a combination of relative technical simplicity and a very high sensitivity (mutation detection rate), approaching 100%. DHPLC can reliably detect single-base mismatches in fragments between 150 and 500 bp, although detection in fragments up to 1,500 bp has been reported. The ability of dHPLC to detect both known and unknown mutations/SNPs, and its' high sensitivity and specificity (reproducibility) has put this technology at the forefront of genetic analysis for a wide variety of diseases.

