Figure 6

Mutation detection of UNC93A in ovarian tumors.A) Mutation detection using P³²-SSCP method. Each exon was amplified with DNAs from tumour sample, its matched control and normal placenta tissue. The variant bands in exon 3 and 4 in the tumour samples and their matched normals compared with normal placenta tissue are indicated by arrows. B) Mutation detection using F-SSCP method. Representative examples are shown for exons 3, 4, and 8. The primers were labelled with fluorescent dye (the forward primers were labelled with HEX, the reverse primers were labelled with FAM). The electrograms are a graphical display of the fluorescent intensity on the y-axis and the mobility along the x-axis. The red peaks are the GeneScan 500 size standard (Perkin Elmer) labelled with TAMRA, which functions as an internal control for each lane. Placenta DNA was used as a normal control. The abnormal peaks in the tumours are indicated by arrows. C) Mutation detection using DHPLC method. PCR was performed using primers flanking exons by 'touchdown' PCR and subjected to DHPLC analysis. The electrograms are a graphical display of the amount of DNA run through the column (intensity peak) on the y-axis and the retention time of the DNA in the column (minute) along the x-axis. The abnormal peaks in exon1 (T59) and exon 5 (T43) are indicated by arrows. PCR products from placenta DNA was used as control.