New Cleavase (R) fragment length polymorphism method improves the mutationdetection assay

Cleavase(R) Fragment Length Polymorphism (CFLP) analysis is a convenient, a ccurate and highly sensitive method for the detection and localization of n ucleic acid mutations. The assay is well suited for high-throughput screeni ng and can be used to detect mutations in known and unknown nucleic acid sa mples. A recent improvement in the CFLP assay termed "temperature ramping" or "ramping" is reported here. This procedural improvement eliminates the n eed for time and temperature optimizations before the actual sample analysi s. In this study, we compare the CFLP ramping procedure to the conventional CFLP optimization procedure and demonstrate equal, and in some cases impro ved, detection of point mutations. With ramping, CFLP reactions are identic al for all DNA fragments analyzed, which allows for increased sample throug hput, decreased assay time and lower overall cost.