DirectBlood Genotyping Kit (#5000) performance with a hydrolysis probe assay
The DirectBlood Genotyping Kit is compatible with a wide range of assays: Here we are detecting a SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hydrolysis probe assay.
Homozygous wild type blood samples will show a strong signal and amplification plot (blue curve) in channel 2 (Cy5) and none or a very low signal
(red curve) in the other channel 1 (ROX).
Heterozygous blood samples will give an intermediate signal and amplification plots with very similar intensities in both channels 1 and 2 (blue and red curve). Homozygous mutant blood samples, channel 1 (ROX) will show a strong signal and amplification plot (red curve) and channel 2 (Cy5) none or a very low signal and amplification plot (blue curve). So the signals are reversed on homozygous mutant samples.
DirectBlood Genotyping Kit performance with a hybridization probe assay
Here we are detecting the same SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hybridization probe assay.
Three blood samples with known genotype were used. All the genotypes can clearly be identified. The blue curve shows the specific wild type (homozygous G;G) peak at 61°C. The red curve shows the specific mutant (homozygous A;A) peak at 53°C. A heterozygous case (A;G) will result in both melting curve peaks, respectively at the same specific temperature for wild type and mutant (orange curve).