Preimplantation diagnosis by whole-genome amplification, PCR amplification, and solid-phase minisequencing of blastomere DNA.

摘要: We have developed a new method for preimplantation diagnosis of inherited diseases. Our procedure the identification point mutations in single cells combines whole-genome amplification using 15-mer random primers (primer extension preamplification, PEP) with locus-specific PCR amplification, followed by detection mutation solid-phase minisequencing. The was evaluated detecting three disease-causing and seven polymorphic nucleotides located on different human chromosomes from granuloma blastomere cells. correct genotype cell identified at 96% nucleotide positions analyzed, showing that representative part genome is amplified during PEP. estimate PEP yielded least 1000 copies genome. quantitative nature minisequencing allowed us to notice preferential one allele occurs heterozygous loci PEP, which potential problem diagnosis.