1. A Novel DHPLC-Based Procedure for the Analysis of COL1A1 and COL1A2 Mutations in Osteogenesis Imperfecta 
Antonella Fuccio, Mariangela Iorio, Felice Amato, Ausilia Elce, Rosaria Ingino, Mirella Filocamo, Giuseppe Castaldo, Francesco Salvatore, Rossella Tomaiuolo 
The Journal of Molecular Diagnostics 
Volume 13, Issue 6, Pages (November 2011)
DOI: /j.jmoldx
Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

2. Figure 1
DHPLC profiles obtained in a patient with OI bearing a mutation in COL1A1 (profile A) and in a control individual bearing a variant in COL1A1 when testing DNA from two cells of buccal mucosa (profile B).
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

3. Figure 2
A: Schematic structure of the pEGFP-C3-Col1A(1-2) minigene constructs and position of the unclassified variants. Green fluorescent protein and COL1A1 or COL1A2 exons are shown as large boxes, and the lines between exons represent the corresponding introns. The thin box indicates the CMV promoter (pCMV), and the black arrows represent the location of the primers used for the splicing pattern analysis. Also shown are alternative predicted splicing donor (d) and acceptor (a) sites, as well as classic splicing donor (D) and acceptor (A) sites. B–D: Splicing pattern analysis of the pEGFP-C3-Col1A(1-2) minigene constructs carrying different genetic variants (M) are compared with their relative wild-type (W) form. The structures of the PCR products are schematically shown to the right of each gel. B: The c _ delTG variant causes complete failure of the normal splicing pattern, which is replaced by a new splicing pattern with the presence of a predominant form that retains intron 11 and a less representative form in which the presence of an alternative donor (d) site into exon 10 causes the alternative splicing of exon 11 and part of exon 10. C and D: The variants c _3046-5dupCT in COL1A1 and c A>T in COL1A2 do not affect splicing.
The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx )
Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions