Validation of the libraries of the serial analysis of gene expression by the application of real-time quantitative polymerase chain reaction

Abstract

To validate and supplement the libraries of serial analysis of gene expression (SAGE) by the application of the real-time quantitative polymerase chain reaction PCR). The primers were designed based on the full sequences of the genes. Nine single matched tags, 6 multiple matched tags, 1 non-matched tag due to the update of the National Center for Biotechnology Information (NCBI) database, and 2 non-matched tags were selected to fulfill the validation of real-time PCR. The genes were all specifically amplified by the primers pairs. The expressions of the single matched tags were identical to those of the SAGE libraries; however, the expressions of only 3 genes of the 6 multi-matched tags were identical to those of the SAGE libraries. The PCR data of the non-matched tag due to the update of the NCBI database were opposite to those of the SAGE libraries. The data did not support the significant difference of the non-matched gene of the SAGE libraries. Real-time PCR is a reliable tool for the validation of high through-put data such as SAGE. The reliability of data depends on the match of the tags of the SAGE libraries.