Figure 1

TSP genotyping by allele-specific PCR of a CT SNP. A. Schematic illustrating the position of TSP primers for allele-specific interrogation of a target region harboring a SNP and the sizes of the expected genotyping products for the two alleles. B. Allele-specific amplification products generated with 0.1 μM LS primers (forward 5'-GGGACATGCATGGTGGCATA and reverse 5'-GAATCTACCACCGCTCCAGCA) and 0.5 μM NLS primers designed to the C allele (forward 5'-GC GGATTGTTGCCCTAC and reverse 5'-CG TGACATTCATTTGTAGTCC). Underlined sequence indicates the non-complementary 5'-tail region. The LS forward primer was positioned 93 bp from the NLS forward primer. The presence of the C allele resulted in the accumulation of a 189 bp allele-specific PCR product (Lanes 3, 8, 9, 12 and 13), and the presence of the T (alternate) allele resulted in the formation of a 282 bp alternate allele PCR product (Lanes 5, 6, 11 and 16). Heterozygous samples resulted in the amplification of both bands, indicating the presence of the two alleles. M represents a pUC19/HpaII DNA size ladder.