3.1 DNA sequencing
A heterozygous DMD splicing mutation, c.2293-1G>C on intron 18, was detected from the proband and his mother but not from the father. It is recognized as a new mutation with WES and Sanger sequencing analysis, being not found in Human Gene Mutation Database (HGMD) (Fig.1).
3.2 Identification of gene inserts in recombinant vectors by colony PCR
The positive colonies were selected and analyzed by colony PCR to determine if the gene inserts of pcMINI- DMD -wt/mut (Fig.2A) or pcMINI-C- DMD -wt/mut were present within the recombinant vertors, respectively (Fig.2B). The gene inserts were confirmed by plasmis sequencing and results were showed in Fig.3A and Fig.4A.