For Research Use Only. Not for use in diagnostic procedures. Others LightMix Kit Alpha1-Antitrypsin Pi S Z RUO LightMix® Kit Alpha1-Antitrypsin Pi*S*Z 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. PID00000650 Detection Kit for Alpha1-Antitrypsin (AAT) Pi*S and Pi*Z mutations, SimpleProbe® format 10 097 639 001 10097639001 LightMix Alpha1-Antitrypsin (PiS, PiZ) LightMix Alpha1-Antitrypsin (PiS, PiZ) 04260159334975 Reagents, kits 40-0576-RUO 64 reactions Not Available false This kit detects the Pi*S and Pi*Z mutations in the Alpha1-Antitrypsin (AAT) gene from human genomic DNA extracts. A 229 bp long PCR fragment containing the G11940A site is analyzed using a LC640 labeled SimpleProbe® (640 channel) which matches the rare 11937A variant. A second PCR fragment with a length of 177 bp includes the T9628A polymorphic site and is analyzed with a 519 labeled SimpleProbe® (530 channel) identical to the 9628A allele. A melting curve analysis is used to determine the genotype. For use on Roche LightCycler® systems. en Using PCR methodology, two fragments of the AAT gene are simultaneously amplified with specific oligonucleotide primers. Fluorescent labeled probes are used to identify the PCR product and to determine the genotype by performing a melting curve analysis.The probe binds to a part of the amplified fragment spanning the mutation site. Any mismatch covered by the probe destabilizes the hybrid. During the melting curve analysis the temperature is slowly increased. The probe melts off at a specific melting temperatures causing the fluorescence to decrease.Pi*Z allele (p.E342K)A 229 bp long PCR fragment containing the G11940A (rs28929474) site is analyzed using a LightCycle® Red 640 labeled SimpleProbe® oligomer which is matching the rare 11937A variant (SNP rs143370956). In the melting curve analysis the 11937A samples display a higher temperature than the wild type allele, while the Pi*Z risk allele 11940A has the lowest melting temperature. Other variants yield intermediate melting temperatures (see chapter 9).Pi*S allele (p.E264V) The second PCR product has a length of 177 bp and includes the T9628A (rs17580) polymorphic site and is analyzed with a 519 labeled SimpleProbe® oligomer identical to the 9628A allele, resulting in a high melting temperature for the Pi*S variant and a lower melting temperature for the wild type allele.Pi*00 allele (gene deletion)In case of a AAT gene deletion there will be no melting peaks visible, but the result cannot be distinguished from missing or low amount of sample DNA or inhibition of amplification or other PCR failures. For samples reporting no melting peaks the test should be repeated and verified with a second method.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').Automated genotyping results must be reviewed by eye for deviating curves and intermediate melting point temperatures. In case that automated typing fails to report consistent genotype results, the genotype must be deducted from the melting temperatures following the criteria described in chapter 8. See also chapter 9 for melting temperatures expected for other variants. en Analytical SensitivityDetection of serial dilutions of several heterozygous human genomic DNAs has revealed that the limit of detection of the present kit is 250 copies (1.5 ng).Analytical SpecificityThe specificity to the target gene and the suitability of the PCR amplification employed in the present test was demonstrated by comparison with the results obtained by direct sequencing. en

LightMix^® Kit Alpha1-Antitrypsin Pi*S*Z

Detection Kit for Alpha1-Antitrypsin (AAT) Pi*S and Pi*Z mutations, SimpleProbe^® format

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LightMix^® Kit Alpha1-Antitrypsin PiSZ

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