title: New Cleavase Fragment Length Polymorphism method improves the mutation detection assay.
abstract: Cleavase Fragment Length Polymorphism (CFLP) analysis is a convenient, accurate and highly sensitive method for the detection and localization of nucleic acid mutations. The assay is well suited for high-throughput screening and can be used to detect mutations in known and unknown nucleic acid samples. A recent improvement in the CFLP assay termed "temperature ramping" or "ramping" is reported here. This procedural improvement eliminates the need for time and temperature optimizations before the actual sample analysis. In this study, we compare the CFLP ramping procedure to the conventional CFLP optimization procedure and demonstrate equal, and in some cases improved, detection of point mutations. With ramping, CFLP reactions are identical for all DNA fragments analyzed, which allows for increased sample throughput, decreased assay time and lower overall cost.
authors.affiliation: Third Wave Technologies, Madison, WI, USA. moldenburg@twt.com
authors.forename: M C
authors.initials: MC
authors.lastname: Oldenburg
authors.forename: M
authors.initials: M
authors.lastname: Siebert
mesh_terms: D001483:Base Sequence; D001709:Biotechnology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004706:Endodeoxyribonucleases; D006801:Humans; D009690:Nucleic Acid Conformation; D011110:Polymorphism, Genetic; D013696:Temperature
chemical_list: D017931:DNA Primers; D004247:DNA; D004706:Endodeoxyribonucleases; C118972:cleavase