For life science research only. Not for use in diagnostic procedures. Others LightMix Kit HLA-B27 LSR LightMix® Kit HLA-B27 100 1377683945183 This product is not a Roche product, Roche Global Customer Support is not offering any Support. For any additional information and support please ONLY contact TIB MolBiol directly. Furthermore we can not guarantee that all available information for this product is up to date. TIBMOLBIOL-0125 Detection of the Human Leukocyte Antigen B27 alleles for use with the Roche Diagnostics LightCycler® Instruments 06 896 537 001 6 896 537 001 06896537001 6896537001 06896537001 LightMix HLA B27 LightMix HLA B27 04260159332261 Reagents, kits 40-0606-64 64 reactions Not Available false Using PCR methodology, a fragment of the HLA-B27 gene is amplified with specific primers. A second fragment from the beta globin gene is amplified as internal control. Both fragments are detected with a specific detection probe internally labeled with SimpleProbe® 519 reagent.SimpleProbes® are fluorescent only when bound to complementary DNA.The probe binds to a part of the amplified fragment. During the melting curve analysis the temperature is slowly increased. The probe melts off at specific melting temperatures causing the fluorescence to decrease.The HLA-B27 probe yields a 8°C higher melting peak than the globin control. HLA-B27 negative samples display only the globin low melting peak, while HLA- B27 positive samples exhibit two melting peaks; sometime the globin amplification is not visible in HLA-B27 positive samples because the assay is devised with a preferential amplification for HLA-B27 target, thus only the high melting peak will be visible.Reading of the genotype results is based on the melting temperatures compared to the supplied standards. If permitted by the instrument software reading of the genotype results can be achieved by the automated genotyping (instrument-dependent: software module 'Melt Curve Genotyping'); in case that automated genotyping fails to report the genotype results, they must be deduct- ed from the melting temperatures following the criteria described in chapter 7. en Analytical SpecificityThe specificity to the target gene and the suitability of the PCR amplification em- ployed in the present test for the detection of the allele variant were demonstrated by sequencing of the generated 133 bp amplicon.Analytical SensitivityDetection of serial dilutions of several heterozygous human genomic DNA has revealed that the limit of detection of the present kit is 350 copies (2 ng). en This kit detects prevalent HLA-B27 alleles from human genomic DNA extracts. A fragment of the HLA-B27 gene is amplified with specific primers. A second fragment from the beta globin gene is amplified as internal control. Both fragments are detected with an internally labeled 519 SimpleProbe® (530 channel). A melting curve analysis is used to determine the genotype. For use on Roche LightCycler® systems. en

LightMix^® Kit HLA-B27

Detection of the Human Leukocyte Antigen B27 alleles for use with the Roche Diagnostics LightCycler^® Instruments

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