Adapter-tagged competitive PCR (ATAC-PCR)--a high-throughput quantitative PCR method for microarray validation

Methods. 2003 Dec;31(4):326-31. doi: 10.1016/s1046-2023(03)00160-9.

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.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Brain Chemistry / genetics
  • Gene Expression Profiling / methods*
  • Mice
  • Microdissection / methods
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligonucleotide Array Sequence Analysis / statistics & numerical data
  • Polymerase Chain Reaction / methods*
  • RNA / analysis

Substances

  • RNA