A high-throughput gene expression analysis technique using competitive PCR and matrix-assisted laser desorption ionization time-of-flight MS.
作者： C. Ding , C. R. Cantor
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摘要: We report here an approach for gene expression analysis by combining competitive PCR and matrix-assisted laser desorption ionization time-of-flight MS. A DNA standard is designed with artificial single nucleotide polymorphism in the of interest. The added to reverse transcription product before PCR. Subsequently, a base extension reaction carried out at position, products are quantified capable relative absolute quantification expression; it extremely sensitive (as few as five copies were quantified) highly reproducible. It also simultaneous both alleles heterozygotes alternatively spliced genes. have incorporated this technique homogeneous Mass Extension system (Sequenom) create high-throughput, automated platform where hundred genes from 20–500 different samples can be accurately per day.
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