Adapter-tagged competitive PCR (ATAC-PCR) – a high-throughput quantitative PCR method for microarray validation |
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Authors: | Sakae Saito Ryo Matoba Kikuya Kato |
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Institution: | a Taisho Laboratory of Functional Genomics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan;b Laboratory of Genetics, National Institute on Aging, National Institutes of Health, Baltimore, MA 21224-6820, USA |
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Abstract: | Adapter-tagged competitive PCR (ATAC-PCR) is an advanced version of competitive quantitative PCR that is characterized by the addition of unique adapters to cDNA derived from each sample RNA. Using multiple adapters, we can accurately measure the relative expression ratios of many samples, with a calibration curve obtained from internal standards included in the same reaction. ATAC-PCR can identify differences in gene expression as small as twofold, even from very small amounts of sample RNA. This technique is suitable for confirming results obtained with cDNA microarrays or differential display, and it can process more than a thousand of genes per day when used in conjunction with a capillary DNA sequencer. |
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Keywords: | Adapter-tagged competitive PCR Gene expression profiling Laser capture microdissection |
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