Fig. 6.

Southern hybridization for the insertion. HaeIII-undigested PCR products were separated in a 1% agarose gel, stained with ethidium bromide (a), and then blotted to a nylon membrane. The membrane was hybridized with biotin-labeled (TAAAA)₅ probe (b), (TGGAA)₅ probe (c), or (TAGAA)₅ probe (d). The insertions in SCA31 patients (lanes 1–3) were clearly detected by (TGGAA)₅ or (TAGAA)₅ probe (c and d) . In patient ID 254 (lane 3), the 3.0-kb insertion (arrow), but not the 4.3-kb insertion (arrowhead), was clearly labeled with (TGGAA)₅ probe (c). In contrast, the 4.3-kb insertion (arrowhead), as well as the insertions in normal controls (lanes 4–6), was more intensively labeled with (TAAAA)₅ probe than the insertions in SCA31 patients (b). The 1.5-kb fragments derived from a normal allele were visualized by (TAAAA)₅ probe (b) because (TAAAA)_n repeats are included in the original genomic sequence. Lanes 1–3 ,SCA31 patients (lane 3, patient ID 254); lanes 4–6, control individuals with the insertion (lane 4: control 1; lane 5: control 2; lane 6: control 3 in Table 1); lanes 7 and 8, control individuals without the insertion