Precision Melt Supermix

Precision melt supermix delivers robust HRM for class I–IV SNPs. Discrimination of A, class I (A to G substitution); B, class II (G to T substitution); C, class III (G to C substitution); and D, class IV (A to T substitution) SNP genotypes from mouse genomic DNA using precision melt supermix. Wild type (), heterozygote (), and homozygous mutant () are shown in the difference plots normalized to wild-type samples. HRM analysis was performed on a CFX384™ real-time PCR detection system and genotypes were automatically assigned by Precision Melt Analysis™ software. Amplification was carried out for 35 cycles. Total run time including melt curve = 150 min. RFU, relative fluorescence units.

Accurate methylation detection with precision melt supermix. A, normalized melt curve; B, difference curve. Mixtures of methylated and unmethylated human genomic DNA of varying ratios were analyzed using HRM on a CFX384™ real-time PCR detection system. Increasing amounts of methylated DNA (, 0%; , 2%; , 5%; , 50%; , 75%; , 95%; and , 100%) were analyzed for methylation of the human RARB2 gene. The genomic region contains 7 CpG sites and is 88 base pairs in length. Total run time including melt curve = 190 min. RFU, relative fluorescence units.

Exceptional stability enables high-throughput genotyping analysis with precision melt supermix. Specific amplification and accurate discrimination of a class IV SNP (84 bp amplicon) from mouse genomic DNA was performed on a CFX384™ real-time PCR detection system either 0 hr (A) or 48 hr (B) after reaction setup. Wild type (), heterozygote (), and homozgous mutant () are shown in the difference plots normalized to wild-type samples. Total run time including melt curve = 150 min. RFU, relative fluorescence units.