© 1993 Oxford University Press
RESEARCH-ARTICLE |
An RT-PCR-SSCP screening strategy for detection of mutations in the gene encoding the 
1 chain of type I collagen: application to four patients with osteogenesis imperfecta
Department of Genetics, University of Leicester Leicester LE1 7RH, UK 1Departments of Pathology and Medicine and the Center for Inherited Disease, University of Washington Seattle, WA 98195, USA
* To whom correspondence should be addressed
Received April 28, 1993; Revised June 10, 1993; Accepted June 10, 1993
A strategy has been developed to allow the screening for mutations In the cDNA of the type I collagen 
1 chain. The method involves polymerase chain reaction amplification of four overlapping segments covering the majority of the sequence coding for 1(1) collagen. Four or five restriction endonuclease digestions are performed for each segment and digested samples subjected to single strand conformation polymorphism analysis. Mutations are finally defined by DNA sequence analysis. The application to the detection of mutations In four patients with lethal and non-lethal forms of osteogenesis Imperfecta is described. In each of these patients, the heterozygous mutation identified resulted In substitution of different glycine residues with serines. In addition to these deleterious mutations, several novel silent variants were characterised. The strategy provides a rapid approach to screening for mutations and polymorphisms In 1(1) collagen cDNA and is of general Interest in scanning large regions of DNA for sequence changes.
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