MutaREAL® HFE from IMMUNDIAGNOSTIK AG
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| Product Name | MutaREAL® HFE |
| Description | To analyze the polymorphism, the variable region of the HFE gene is amplified using realtime PCR (e.g. the LightCycler® system). Genomic DNA serves as a template for thisamplification. Using this amplicon a melting curve analysis is performed that incorporatesspecific FRET-hybridisation probes. This melting curve analysis facilitates the distinctidentification of the HFE allele.For the real time PCR reaction in the LightCycler® capillary two primer pair are used thatflank the variable region in the HFE gene and produces a fragment with a length of 291bp for H63D and S65C or 365 bp for C282Y. In addition two probes (fluorescentlabeled sequence specific oligonucleotides) are incorporated into the PCR reaction mix.Both bind the target DNA including the polymorphism in close proximity in a regionbetween the primer pair. Upon this binding a fluorescent signal is generated that isdetected by the optical unit of the real time PCR instrument (e.g. the LightCycler®).For the genotyping analysis a melting curve is recorded after the amplification reaction.This is achieved by constantly increasing the temperature. Perfect homologous probesrequire a higher temperature to be melted as those having a mismatch (the sequence ofthe reporter probe and target DNA matches to 100 %). The sequence of the reporterprobe matches the mutated DNA for H63D and the wildtype DNA for C282Y perfectly.Hence the melting curve of the mismatch (wildtype for H63D and mutation for C282Y)occurs earlier needing a lower temperature to achieve the maximum as the mutationallele.4 2017 Immundiagnostik AG / Version 1.2In very rare cases of heterozygous mutations H63D and S65C no wild-type peak will bedetected, due to the probe used and to the proximity of these two mutations, only theexact matching probes can bind. The result looks as a (not yet described) homozygousdouble mutation H63D + S65C. |
| Size | 96 tests |
| Concentration | n/a |
| Applications | n/a |
| Other Names | HFE |
| Gene, Accession, CAS # | n/a |
| Catalog # | KF291096 |
| Price | please inquire |
| Order / More Info | MutaREAL® HFE from IMMUNDIAGNOSTIK AG |
| Product Specific References | n/a |
IMMUNDIAGNOSTIK AG
IMMUNDIAGNOSTIK AG
Stubenwald Allee 8a
64625 Bensheim GERMANY
P: 49-6251701900
F: 49-6251849430
info@immundiagnostik.com
http://www.immundiagnostik.com
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