MUTATION SCREENING BY A COMBINATION OF BIOTIN-SSCP AND DIRECT SEQUENCING

We have developed a mutation detection strategy that combines single s trand conformational polymorphism (SSCP) analysis of one strand of a d ouble-stranded amplification product with direct sequencing of the oth er. Using this strategy, which we find economical of both time and res ources, we have identified a G to A transition, which substitutes a se rine for glycine residue at position 862 in the major helix of the al chain of Type I collagen. We use this mutation, which causes a lethal form of osteogenesis imperfecta, to illustrate the technique.