Biotechnology Letters, Vol.25, No.19, 1613-1618, 2003
Detection of known mutations in hypertrophic cardiomyopathy using oligonucleotide microarrays assisted by improved base stacking hybridization
With the assistance of improved base stacking hybridization, a low-density microarray, containing 12 capture probes, was used to identify 7 known hypertrophic cardiomyopathy-related mutations in the gene of MYH7 (beta-myosin heavy chain). The hybridization targets, amplified from 11 plasmids containing wild type or mutation sequences of MYH7 and healthy genomic DNA, were prepared by single-step fluorescence labeled asymmetric PCR. Six single base substitutions and a trinucleotide deletion were identified unambiguously, and the average discrimination ratio (Q(mut)) for artificial heterozygous samples was as high as 16.2.
Keywords:base stacking hybridization;beta-myosin heavy chain;hypertrophic cardiomyopathy;microarray;mutation detection