High throughput methods and kits for single nucleotide polymorphism (SNP)
genotyping are provided. The methods involve utilizing nested PCR
amplification reactions which produce sequencible and ligatible
structures. An outer PCR primer set amplifies the SNP, and an inner PCR
primer set amplifies a portion of the DNA amplified by the outer primer
set, but does not amplify the SNP itself. The inner and outer primers may
reaction include non-target common domain sequences, and the inner primer
common domain sequences may comprise digestion restriction endonuclease
recognition sites. The design of the inner primer set allows precise
tailoring of the sequencible and ligatible structures with respect to
length and base composition.