Amplification of NB1-genomic fragments by PCR and exon organization of NB1 cDNA.

(A) Primers were numbered according to the NB1 cDNA reference sequence.12 5.8 kbp fragment: FP1 (409-426) and reverse primer RP1 (859-879) (lanes 2-4). 1.3 kbp fragment: FP2 (329-348) and RP 2 (416-436) (lanes 7-9). Lanes 4 and 9: NB1⁻ patient 1 and patient 2, respectively. Lanes 3 and 8: NB1-expressing healthy persons. Lanes 1 and 6: molecular weight standards. Lanes 2 and 7: negative controls. (B) Genomic DNA from NB1⁺ and NB1⁻ patients was amplified by PCR and sequenced. Exon–intron borders were determined according to the NB1 cDNA reference sequence12 and dinucleotides gt/ag at the intron termini.