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Rapid Genotyping (rapid + genotyping)
Selected AbstractsSimultaneous genotyping to detect myostatin gene polymorphism in beef cattle breedsJOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 6 2002M. E. Miranda Summary The myostatin gene codes for a growth factor involved in muscle development, and polymorphism in this gene can have important economic consequences. Nine mutations affecting the amino-acidic sequence have already been described, six of which are disruptive, inactivating the protein and causing bovine muscular hypertrophy. As the number of known mutations grows, it is necessary to develop a simple, routinely usable technique able to screen individuals in all populations. The oligonucleotide ligation assay (OLA) is proposed here for the rapid genotyping of the nine mutations known affecting the coding sequence in the main breeds of beef cattle. This technique showed its ability to reveal the genotype of individuals being a good tool to determine the frequency of each mutation in a population. The procedure is very flexible as new mutations can be added and removed at any time. Depending on the genotype of each individual, the technique allows breeders to make quick decisions on matings and general selection tendencies. Zusammenfassung Simultane Genotypisierung von Polymorphismen im Myostatin-Gen in Fleischrinderrassen Das Myostatin-Gen kodiert für einen Wachstumsfaktor, der in die Muskelentwicklung eingebunden ist und Polymorphismen in diesem Gen können daher wichtige ökonomische Konsequenzen haben. Bisher wurden neun Mutationen, die Auswirkungen auf die Aminosäuresequenz haben, beschrieben. Sechs davon inaktivieren das Protein und verursachen bovine muskuläre Hypertrophie. Da die Anzahl der bekannten Mutationen in diesem Gen steigt, ist es notwendig, eine einfache, in der Routine einsetzbare Methode zu entwickeln, um Individuen in allen Populationen untersuchen zu können. Zur schnellen Genotypisierung der neun bekannten Mutationen, welche die kodierende Sequenz in den Hauptfleischrinderrassen betreffen, wird hier der Oligo-Ligationsassay (OLA) vorgeschlagen. Durch diese Technik ist es möglich, den Genotyp jedes Individuums und die Frequenz jeder einzelnen Mutation in der Population festzustellen. Die Prozedur ist sehr flexibel, da zu jedem Zeitpunkt neue Mutationen hinzugefügt bzw. weggelassen werden können. Diese Methode erlaubt dem Züchter, in Abhängigkeit vom Genotyp jedes Individiums schnelle Entscheidungen über die Anpaarung und die allgemeine Selektionsrichtung zu treffen. [source] A rapid real-time PCR assay for determination of hepatitis C virus genotypes 1, 2 and 3aJOURNAL OF VIRAL HEPATITIS, Issue 4 2006A. Moghaddam Summary., The genotypes of hepatitis C virus (HCV) in serum of patients have been described as independent predictors of success of antiviral therapy. Therefore, different antiviral regimens have been proposed depending on the infecting HCV genotype. HCV strain is usually determined by polymerase chain reaction (PCR) amplification of genome followed by sequencing or by line-probe assays. We report a new one step real-time PCR assay for genotyping of HCV strains that are prevalent in patients in Norway. HCV types 1, 2 and 3a were genotyped unambiguously in 37 patient serum samples with 100% correlation to genotyping by nucleotide sequence analysis and line-probe assays. Genotyping could also be confirmed against an HCV genotype panel from the National Institute for Biological Standards and Control. This assay does not require manipulation of amplified PCR products, it involves very little hands on and analysis time. This assay can be used for rapid genotyping of HCV-RNA in infected patients to aid physicians decide suitability of patients for treatment and subsequent length of treatment. [source] Rapid DNA extraction and PCR-sexing of mouse embryosMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 2 2001Peter J. McClive Abstract We have devised a PCR-based sexing method that is quick, simple, and highly reproducible. DNA is first extracted from embryonic mouse yolk sac via a 15 min, two-step incubation procedure utilizing PCR-compatible proteinase K buffer. Without any further manipulation the lysate is subjected to 30 cycles of PCR, optimized to run in less than 1 hr. The reaction includes multiplexed primer pairs for Sry and Myog (myogenin) that generate a male specific band of 441 bp and an internal control band of 245 bp, respectively. This robust method is used routinely in our laboratory and gives rapid genotyping results with 98% reliability and 100% accuracy. Mol. Reprod. Dev. 60: 225,226, 2001. © 2001 Wiley-Liss, Inc. [source] Flanking genomic region of Tyr::Cre mice, rapid genotyping for homozygous micePIGMENT CELL & MELANOMA RESEARCH, Issue 4 2007Sophie Colombo No abstract is available for this article. [source] |