Bi-directional Dideoxy Fingerprinting (Bi-ddF): Rapid and Efficient Screening for Mutations in the Big Blue~(TM) Trans-genie Mouse Mutation Detection System

摘要

Single-stranded conformation polymorphism (SSCP) is currently the most widely used method for screening for mutations. SSCP does not detect all sequence changes, and its sensitivity depends on the sequence and size of the segment screened. For a sequence of about 200 bp, the observed sensitivities are often in the range of 60%-90%. For larger segments, the sensitivity is decreased. Dideoxy fingerprinting (cldF) is performed by elec-trophoresing one lane of a Sanger dideoxy termination reaction through a non-denaturing gel. ddF can screen a 250-bp segment with essentially 100% sensitivity.