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Renewable Standard Reference Material for the Detection of TP53 Mutations

Published

Author(s)

C D. O'Connell, L A. Tully, J E. Devaney, M A. Marino, J P. Jakupciak, Donald H. Atha

Abstract

It is important to have standards for use in mutation detection systems to monitor the sensitivity of mutation detection and the possible introduction of errors during amplification and separation procedures. We have developed 12 plasmid clones containing a 2.0 kilobase region of the TP3 gene including exons 5-9. Eleven of these clones contain a single mutation within the mutational hot spots of the TP53 gene, the twelfth is wild type in this region of the gene. We have analyzed the single point mutations by capillary electrophoresis single strand conformational polymorphism (CE-SSCP), denaturing gradient gel electrophoresis (DGGE), and denaturing high performance liquid chromatography (DHPLC), as well as full sequence analysis of both DNA strands of the cloned material. Results were compared for the three detection systems. The SRM reference panel was successfully implemented to validate mutation detection across these different technology platforms.
Volume
7;2
Conference Dates
February 1, 2003
Conference Location
Undefined
Conference Title
Molecular Diagnostics

Keywords

CD-SSCP, DGGE, DHPLC, DNA standards, heteroduplex analysis, single-strand convormation polymorphism, SSCP, TP53 mutation detection

Citation

O'Connell, C. , Tully, L. , Devaney, J. , Marino, M. , Jakupciak, J. and Atha, D. (2021), Renewable Standard Reference Material for the Detection of TP53 Mutations, Molecular Diagnostics, Undefined (Accessed July 7, 2022)
Created October 12, 2021