- DirectBlood Genotyping PCR Kit is a novelty on the market. It allows genotyping in real-time and without any previous DNA isolation. You simply save time and money: directly use EDTA blood without any extraction steps.
The DirectBlood Genotyping PCR Kit is optimized to function with a wide range of different SNPs to detect possible nucleotide variations. It has been successfully tested with a variety of hydrolysis probes and hybridization probes.The setup of a genotyping experiment can not be simpler anymore: Mix primers&probes with the Genotying mastermix, add the respective blood sample and start real-time PCR, this is it.
It is even more simple: store the DirectBlood Genotyping PCR Kit at room-temperature. A cool chain is not needed anymore. The included master mix is freeze-dried and dissolves within a few seconds after addition of the rehydration buffer.Additionally, we have already established SNP-assays for you. In case, you can not find your respective SNPs, please provide us with the rs-number of the SNP-assays your are interested-in and you will receive an individual quotation from us.For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
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).
