Technical note: A novel method for routine genotyping of the G allele of β-casein (CSN2) and T allele of κ-casein (CSN3) in a sheep population using LightCycler

The goal of this work was to develop a protocol for rapid genotyping of A and G variants at the CSN2 locus and genotyping of T and C variants at the CSN3 locus in sheep breeds (Sumava and Valachian) by means of PCR and LightCycler analysis. The LightCycler technique combines rapid and efficient in vitro amplification of DNA in glass capillaries with melting curve analysis based on fluorescence resonance energy transfer for the sensitive detection of point mutation. The A variant had a greater frequency (Sumava, 0.778; Valachian, 0.835) than did variant G (Sumava, 0.222; Valachian, 0.165) in both sheep breeds. The CSN3 locus was found to be monomorphic, with no polymorphism identified in either population.