LightMix® Kit MTHFR C677T -Research Use Only
Kit with reagents for the detection of the human MTHFR C677T polymorphism using the Roche Diagnostics LightCycler® Instrument.
Methylenetetrahydrofolate reductase (MTHFR) catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a co-substrate for the remethylation of homocysteine to methionine. The absence of 5-methyltetrahydrofolate induces an increase of homocysteine in plasma. Several polymorphisms of MTHFR are known. The best studied substitution C677T is associated with an increased concentration of homocysteine in plasma due to a reduced activity of the enzyme. This is based on a conversion of the amino acid alanine to valine. The LightMix® Kit MTHFR C677T provides a fast, easy and accurate system to identify the genotype of this target in a nucleic acid extract. This LightMix® Kit is tested on the LightCycler® 2.0 Instrument with Roche Diagnostics ‘LightCycler® FastStart DNA Master HybProbe’.
A 233 bp fragment of the human MTHFR gene is amplified with specific primers. The resulting PCR fragments are analyzed with hybridization probes labeled with LightCycler® Red 640 (detected in channel 640). The genotype is identified by running a melting curve with specific melting points (Tm). The wildtype MTHFR C677 DNA exhibits a Tm of 63.0°C in channel 640. The muta nt MTHFR 677T exhibits a Tm of 54.5°C in channel 640. The supplied control DNA allows for the accurate comparison with unknown samples.
PCR results are obtained within 45 minutes (45 cycles and melting curve) with the LightCycler® 2.0 Instrument.
- These reagents detect 1 ng of human genomic DNA using the Roche ‘LightCycler® FastStart DNA
Master HybProbe’ with the LightCycler® 2.0 Instrument.
- The measuring range of the assay is 1 ng to 100 ng of human genomic DNA using the Roche ‘LightCycler® FastStart DNA Master HybProbe’ with the LightCycler® 2.0 Instrument.
Storage and Stability
- Lyophilized reagents are stable for at least 3 months after shipment when stored protected from light at room temperature (18-25°C).
- Do not freeze lyophilized reagents.
- Dissolved reagents are stable for at least 5 days when stored protected from light and refrigerated (4°C).