Mutation detection methods

Analysis of Genetic Variability by PCR-coupled Mutation Detection Methods... 

Stabilized, and voltammetric technique is stable. When compared with conventional methods, it can be observed that electrochemical techniques are also capable of sequence specificity and allele specificity, time-consuming, and highly sensitive. Further research-based mutation detection methods are under progress in our laboratory. 

Limbic system detect a gene mutation. The method uses the coupling of two adjacent synthetic oligonucleotides aligned on the template of the target deoxyribonucleic acid (DNA). Area of the brain associated with small, involuntary functions, emotions and behaviour. Comprises the hypothalamus, parahippocampus, olfactory lobe, dentate gyrus, amygdala, anterior thalamus, fornix and stria terminalis. 

Levin D-E, Hollstein M, Christman M-F, Schwiers E-A and Ames B-N (1982) A new Salmonella tester strain with A.T base pairs at the site of mutation detects oxidative mutagens. Proc Nad Acad Sci USA 79 7445-7449 Maron D-M, Ames B-N (1983) Revised methods for the Salmonella mutagenicity test. Mutation Res 113 173-215 McCann J, Springarn N-E, Kobory J and Ames B-N (1975) Detection of carcinogens as mutagens bacterial tester strains with R factor plasmids. Proc Nat Acad Sci USA 72 979-983... 

Overview of Ligase-based Methods for DNA Mutation Detection. 172... 

In the last decade, a large amount of effort has been devoted to the development of detection methods which are compatible for use in combination with DNA sensor array platforms and are sufficiently sensitive to detect single point mutations using minute amounts of sample [164,166]. Electrochemical detection methods are a particularly interesting alternative to the methods described above due to their simplicity and low cost. 

A commonly used approach, known as the competitive PCR method, exploits the competition and plateau features of PCR to quantitate the expression of the mRNA of interest (15,16). The reaction incorporates an RNA or DNA PCR template which is included with the sample and amplified along with the RNA of interest. The most frequently used alternate methods include the use of competitive RNA or DNA PCR templates that are different in size, contain a recognizable mutation detectable because of gel electrophoresis separation properties, or contain a restriction site to allow digestion following PCR. Typically, in these approaches, serial dilutions of a known concentration of the competitor RNA or DNA are added to tubes containing equal, but unknown quantities of the mRNA of interest. Subsequently, RT is performed, followed by PCR amplification. The differing PCR products are separated by gel electrophoresis, and the concentration in the unknown sample is obtained at the dilution where the product of the unknown sample is equal to the product of the competitor. 

S. (1996) Mutation detection by highly sensitive methods indicates that p53 gene mutations in breast cancer can have important prognostic value. Proc. Natl. Acad. Sci. USA 93, 1093-1096. 

Mutation detection by DNA analysis is specific and much more sensitive than biochemical methods. ° It is therefore fast replacing other methods, particularly since it has the additional advantage of enabling asymptomatic disease to be excluded with certainty. However, it depends on prior identification of a disease-specific mutation in the family under investigation. In the 5% or so of families in which a mutation cannot... 

Dolinsky LCB. Denaturing Gradient Gel Electrophoresis (DGGE) for mutation detection in Duchenne muscular dystrophy (DMD). Methods Molec Biol 2003 217 165-75. 

Two genotyping methods are reviewed in detail because they are frequently used in laboratories in the United States and, more importantly, they have peer-reviewed articles describing their characteristics. The Ampli-Chip CYP 450 Test [25,26] was cleared for clinical use by the FDA at the beginning of 2006 the Luminex Tag-It Mutation Detection Kit [29 31] is currently approved for research use only. A prior article [25] described other CYP genotyping methods mainly used for research purposes and the commercial laboratories in the United States providing CYP genotyping. 

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