A point mutation in the splice donor site of intron 7 in the as2-casein gene of the River buffalo results in an allele-specific exon skipping

The CSN1S2 cDNA of 10 unrelated Mediterranean River Buffaloes reared in Southern Italy was amplified by RT-PCR, while the region from the 6th to the 8th exon of the CSN1S2 gene was amplified from genomic template. All amplicons were sequenced twice and in both directions. Fiftythree individuals randomly chosen from four breeding herds were genotyped for an AluI-restriction fragment length polymorphism (RFLP). Primer sequences and PCR conditions are given in Tables S1 & S2. Three individual milk samples from buffaloes with different genotypes at CSN1S2 were analysed by reverse-phase-high pressure liquid chromatography (RP-HPLC).