Analysis of multiple single nucleotide polymorphisms closely positioned in the ovine PRNPgene using linear fluorescent probes and melting curve analysis

Table 3 Peak area ratios and imbalanced melting peaks

Heterozygote	Mean area ratio	SD	N	Lower limit	Upper limit
136 A/V	0.83	0.07	238	0.62	1.04
154 R/H	0.61	0.06	396	0.43	0.79
171 R/Q	0.97	0.08	1516	0.73	1.21
171 R/H	0.52	0.08	153	0.28	0.76
171 Q/H	0.75	0.06	46	0.57	0.93

4560 purified DNA samples were employed to determine the acceptance criteria for peak area ratios. The peak area ratios of heterozygous samples were calculated by dividing the area under the mismatched melting peak by the area under the matched melting peak. N is the number of heterozygous samples and SD is the standard deviation. Acceptance criteria for normal balanced heterozygotes were determined as the mean area ratio ± 3 standard deviations. Samples exhibiting imbalanced melting peaks generated area ratios outside of defined limits.

ISSN: 1471-2334