Fig. 2.
Identification of the maternal mutation at the splice donor site in intron 2. (A) Sequence (sense) of the exon 2/intron 2 junction in amplified genomic DNA. Genomic fragments containing exon 2 were amplified from a control (left) and the patient (right) and sequenced directly. A mutation at the splice donor site in intron 2 (G+1 → A) is indicated by arrows. (B) Detection of the splice site mutation by the BspMI digestion. Amplified genomic fragments (690 bp) from intron 1 (bp 14,901) to intron 2 (base pair 15,590) was digested with BspMI, electrophoresed in 2.0% agarose gel, and stained with ethidium bromide. The fragment has twoBspMI recognition sites at bp 15,282 and 15,344, predicting 62-, 246-, and 382-bp fragments in the control lane. Loss of theBspMI site (base pair 15,282) by the mutation results in the undigested fragment (308 bp). Lane 1, control; lane 2, father; lane 3, mother; and lane 4, patient. BspMI digestion shows the patient and his mother were heterozygous for the splice site mutation.

Identification of the maternal mutation at the splice donor site in intron 2. (A) Sequence (sense) of the exon 2/intron 2 junction in amplified genomic DNA. Genomic fragments containing exon 2 were amplified from a control (left) and the patient (right) and sequenced directly. A mutation at the splice donor site in intron 2 (G+1 → A) is indicated by arrows. (B) Detection of the splice site mutation by the BspMI digestion. Amplified genomic fragments (690 bp) from intron 1 (bp 14,901) to intron 2 (base pair 15,590) was digested with BspMI, electrophoresed in 2.0% agarose gel, and stained with ethidium bromide. The fragment has twoBspMI recognition sites at bp 15,282 and 15,344, predicting 62-, 246-, and 382-bp fragments in the control lane. Loss of theBspMI site (base pair 15,282) by the mutation results in the undigested fragment (308 bp). Lane 1, control; lane 2, father; lane 3, mother; and lane 4, patient. BspMI digestion shows the patient and his mother were heterozygous for the splice site mutation.

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