Bediaga Naiara G, Alfonso-Sánchez Miguel A, de Renobales Mertxe, Rocandio Ana M, Arroyo Marta, de Pancorbo Marian M
Servicio de Genómica, Banco de ADN, Facultad de Farmacia, Universidad del País Vasco, 01006 Vitoria, Spain.
Anal Biochem. 2008 Jul 15;378(2):221-3. doi: 10.1016/j.ab.2008.04.010. Epub 2008 Apr 9.
GSTT1 and GSTM1 genes possess an inherited deletion associated with a lack of enzyme activity. The heterozygous condition of this deletion is difficult to determine in low-quality DNA with existing PCR protocols. We designed and validated a multiplex real-time PCR assay by adapting the DeltaDeltaCt relative quantification method for the analysis of GSTT1 and GSTM1 markers to accurately differentiate the three genotypes ( *1/1, *1/0, and *0/0) in degraded DNA from formalin-fixed paraffin-embedded tissue. Gene copy number values obtained provide for unambiguous homozygous and heterozygous differentiation. The efficacy shown by the PCR assay endorses its usefulness for complete genotyping of glutathione S-transferases in archival tissues.
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