Figure 2.

A) The cDNA from patient 138 (carrier of c.583C>G in exon 3 of SPAST) was synthesised from leukocytes mRNA. The sequencing of the fragment amplified with primers that matched exons 2 and 7 showed the deletion of the last 4 nucleotides of exon 3. B) The cDNA from patient 212 (X617Glu) was amplified with primers for exons 11 and 17 of SPAST, and the sequence showed similar electropherogram peaks for the two alleles. C) The cDNA of patient 322 was amplified with primers for exons 7 and 17 of SPAST, and the sequence showed the effect on splicing of the c.1414-1G>C mutation. D) MLPA of the SPAST gene in patient 199, showing the exons 10-16 deletion. The cDNA was amplified with primers that matched exons 9 and 17, and two PCR fragments were obtained. Sequencing of the smaller fragment confirmed the absence of exons 10 to 16. P = patient (cDNA); M = Patient's mother's (cDNA); C = Control (cDNA); G = Genomic DNA