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  • Articles  (2)
  • DNA p53 mutation detection  (1)
  • Forensic  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Detection of p53 point mutations by single strand conformation polymorphism: Analysis by capillary electrophoresis (1998)
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 172-179 
    Details
    ISSN: 0173-0835
    Keywords: DNA p53 mutation detection ; Capillary electrophoresis ; Slab gel electrophoresis ; Single strand conformation polymorphism ; Temperature dependence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have analyzed five p53 single point mutations by single strand conformation polymorphism using capillary electrohoresis (CE-SSCP) and have compared these measurements to measurements obtained by slab gel electrophoresis (SG-SSCP). PCR primers were used for amplification of specific exons for mutation detection. 5′ Primers were labeled with FAM (5-carboxyfluorescein) and 3′ primers were labeled with JOE (2′,7′-dimethoxy-4′,5′-dichloro-6-carboxyfluorescein). CE-SSCP was performed using the Perkin Elmer ABI PRISM™ 310 Genetic Analyzer with GeneScan™ Software and the Beckman P/ACE™ 5510 CE equipped for laser-induced fluorescence detection. Although the shifts in migration times for the p53 mutations relative to the corresponding wild-type strands could be successfully detected by either SG or CE analysis, the individual electrophoresis run times were about tenfold faster and more automated with capillary electrophoresis. The CE-SSCP measurements were performed at temperatures ranging from 10 to 60°C on a prototype instrument. For mutations measured at ambient temperature (25°C), characteristic shift in direction and magnitude were observed in the migration times to both strands of all mutations relative to the wild type. This demonstrated the ability of CE at ambient temperature to resolve these mutations. However, the magnitute and direction of shifts in migration time varied with temperature in a discrete pattern for each mutation and resulted in a temperature-specific profile for each mutation. This demonstrated that extended temperature control will be an important advantage in resolving single point mutations by CE-SSCP. In addition, by using CE, discrete intra-strand isoforms could be easily observed at different temperatures. The combination of mutation-specific temperature profiling and analysis of isoforms by CE-SSCP should be of help to the diagnostic community in the detection of genetic mutations.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150190207
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  • 2
    Electronic Resource
    Electronic Resource
    Genotyping of forensic short tandem repeat (STR) systems based on sizing precision in a capillary electrophoresis instrument (1998)
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 86-93 
    Details
    ISSN: 0173-0835
    Keywords: Capillary electrophoresis ; Forensic ; Short tandem repeats ; Genotyping ; AmpFlSTR ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Automated fluorescence analysis of polymerase chain reaction (PCR)-amplified short tandem repeat (STR) systems by capillary electrophoresis (CE) is becoming an established tool both in forensic casework and in the implementation of both state and national convicted offender DNA databases. A new capillary electrophoresis instrument, the ABI Prism 310 Genetic Analyzer, along with the Performance Optimized Polymer 4 (POP-4) provides an automated and precise method for simultaneously analyzing ten flourescently labeled STR loci from a single PCR amplification kit, which provides a power of discrimination of approximately one in five billion from a single PCR amplification. Data are presented on sizing precision, sizing accuracy, and resolution for the STR loci in the AmpFlSTR Profiler™ kit. Sizing accuracy is highly dependent on the electrophoresis system, and therefore the reporting of alleles based on the nucleotide size obtained from an electrophoresis system is not recommended for forensic work. The precision of the 310 capillary electrophoresis system, coupled with software developed for automated genotyping of alleles based on the use of an allelie ladder, allows for accurate genotyping of STR loci. Sizing precision of ≤ 0.16 nucleotide standard deviation was obtained with this system, thus allowing for accurate genotyping of length variants that differ in length by a single nucleotide.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150190116
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